Inhibitors of the interaction between MDM2 and P53

ABSTRACT

The present invention provides compounds of formula (I), their use as an inhibitor of a p53-MDM2 interaction as well as pharmaceutical compositions comprising said compounds of formula (I) 
                         
wherein n, m, p, s, t, R 1 , R 2 , R 3 , R 4 , R 5 , R 6 , R 7 , X, Y, Q and Z have defined meanings.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a divisional of U.S. application Ser. No. 11/575,552filed on Mar. 19, 2007 now U.S. Pat. No. 7,834,016, which is theNational Stage application under 35 U.S.C. §371 of PCT/EP2005/054604,filed Sep. 16, 2005, which claims priority from EPO Patent ApplicationNo. 04077630.4, filed Sep. 22, 2004, and U.S. provisional applicationNo. 60/613,902, filed Sep. 28, 2004, the entire disclosures of which arehereby incorporated in their entirety.

FIELD OF THE INVENTION

The present invention relates to compounds and compositions containingsaid compounds acting as inhibitors of the interaction between MDM2 andp53. Moreover, the present invention provides processes for thepreparation of the disclosed inhibitors, compositions comprising themand methods of using them, for instance as a medicine.

p53 is a tumour suppressor protein which plays a pivotal role in theregulation of the balance between cell proliferation and cell growtharrest/apoptosis. Under normal conditions the half life of p53 is veryshort and consequently the level of p53 in cells is low. However, inresponse to cellular DNA damage or cellular stress (e.g. oncogeneactivation, telomere erosion, hypoxia), levels of p53 increase. Thisincrease in p53 levels leads to the activation of the transcription of anumber of genes which drives the cell into either growth arrest or intothe processes of apoptosis. Thus, an important function of p53 is toprevent the uncontrolled proliferation of damaged cells and thus protectthe organism from the development of cancer.

MDM2 is a key negative regulator of p53 function. It forms a negativeautoregulatory loop by binding to the amino terminal transactivationdomain of p53 and thus MDM2 both inhibits the ability of p53 to activatetranscription and targets p53 for proteolytic degradation. Under normalconditions this regulatory loop is responsible for maintaining the lowlevels of p53. However, in tumours with wild-type p53, the equilibriumconcentration of active p53 can be increased by antagonising theinteraction between MDM2 and p53. This will result in restoration of thep53-mediated pro-apoptotic and anti-proliferative effects in such tumourcells.

MDM2 is a cellular proto-oncogene. Over-expression of MDM2 has beenobserved in a range of cancers. MDM2 is over-expressed in a variety oftumours due to gene amplification or increased transcription ortranslation. The mechanism by which MDM2 amplification promotestumorigenesis is at least in part related to its interaction with p53.In cells over-expressing MDM2 the protective function of p53 is blockedand thus cells are unable to respond to DNA damage or cellular stress byincreasing p53 levels, leading to cell growth arrest and/or apoptosis.Thus after DNA damage and/or cellular stress, cells over-expressing MDM2are free to continue to proliferate and assume a tumorigenic phenotype.Under these conditions disruption of the interaction of p53 and MDM2would release the p53 and thus allow normal signals of growth arrestand/or apoptosis to function.

MDM2 may also have separate functions in addition to inhibition of p53.For example, it has been shown that MDM2 interacts directly with thepRb-regulated transcription factor E2F1/DP1. This interaction could becrucial for the p53-independent oncogenic activities of MDM2. A domainof E2F1 shows striking similarity to the MDM2-binding domain of p53.Since the interactions of MDM2 with both p53 and E2F1 locate to the samebinding site on MDM2, it can be expected that MDM2/p53 antagonists willnot only activate cellular p53 but also modulate E2F1 activities, whichare commonly deregulated in tumour cells.

Also the therapeutic effectiveness of DNA damaging agents currently used(chemotherapy and radiotherapy), may be limited through the negativeregulation of p53 by MDM2. Thus if the MDM2 feed-back inhibition of p53is interrupted, an increase in functional p53 levels will increase thetherapeutic effectiveness of such agents by restoring the wild-type p53function that leads to apoptosis and/or reversing of p53-associated drugresistance. It was demonstrated that combining MDM2 inhibition andDNA-damaging treatments in vivo led to synergistic anti-tumour effects(Vousden K. H., Cell, Vol. 103, 691-694, 2000).

Thus disruption of the interaction of MDM2 and p53 offers an approachfor therapeutic intervention in tumours with wild-type p53, might evenexhibit anti-proliferative effects in tumour cells that are devoid offunctional p53 and furthermore can sensitise tumorigenic cells forchemotherapy and radiotherapy.

BACKGROUND OF THE INVENTION

JP 11130750, published on 18 May 1999, describes amongst others,substituted phenylaminocarbonylindolyl derivatives as 5-HT receptorantagonists.

EP1129074, published on 18 May 2000, describes anthranilic acid amidesas inhibitors of vascular endothelial growth factor receptors (VEGFR)and useful in the treatment of angiogenic disorders.

EP1317443, published on 21 Mar. 2002, discloses tricyclic tert-aminederivatives, useful as chemokine receptor CXCR4 or CCR5 modulators fortreating human immunodeficiency virus and feline immunodeficiency virus.

EP1379239, published on 10 Oct. 2002, disclosesN-(2-arylethyl)benzylamines as antagonists of the 5-HT₆ receptor. Morein particular

6-chloro-N-[[3-(4-pyridinylamino)phenyl]methyl]-1H-indole-3-ethanamine,

N-[[3-(4-pyridinylamino)phenyl]methyl]-1H-indole-3-ethanamine, and

5-methoxy-N-[[3-(4-pyridinylamino)phenyl]methyl]-1H-indole-3-ethanamine,

are described.

WO00/15357, published on 23 Mar. 2000, provides piperazine-4-phenylderivatives as inhibitors of the interaction between MDM2 and p53.EP1137418, published on 8 Jun. 2000, provides tricyclic compounds forrestoring conformational stability of a protein of the p53 family.

WO03/041715, published on 22 May 2003, describes substituted1,4-benzodiazepines and the uses thereof as inhibitors of the MDM2-p53interactions.

WO03/51359, published on 26 Jun. 2003, providescis-2,4,5-triphenyl-imidazolones that inhibit the interaction of MDM2protein with p53-like peptides and have antiproliferative activity.

WO04/05278, published on 15 Jan. 2004, discloses bisarylsulfonamidecompounds that bind to MDM2 and can be used in cancer therapy.

There continues to be a need for effective and potent small moleculesthat inhibit the interactions between MDM2 and p53.

The compounds of the present invention differs from the prior art instructure, in their pharmacological activity and/or in pharmacologicalpotency.

DESCRIPTION OF THE INVENTION

The present invention provides compounds, compositions for, and methodsof, inhibiting the interactions between MDM2 and p53 for treatingcancer. Furthermore the compounds and compositions of the presentinvention are useful in enhancing the effectiveness of chemotherapy andradiotherapy.

This invention concerns compounds of formula (I)

A compound of formula (I),

-   a N-oxide form, an addition salt or a stereochemically isomeric form    thereof, wherein-   m is 0, 1, or 2 and when m is 0 then a direct bond is intended;-   n is 0, 1, 2, or 3 and when n is 0 then a direct bond is intended;-   p is 0, or 1 and when p is 0 then a direct bond is intended;-   s is 0, or 1 and when s is 0 then a direct bond is intended;-   t is 0 or 1 and when t is 0 then a direct bond is intended;-   X is C(═O) or CHR⁸; wherein    -   R⁸ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, —C(═O)—NR¹⁷R¹⁸,        hydroxycarbonyl, arylC₁₋₆alkyloxycarbonyl, heteroaryl,        heteroarylcarbonyl, heteroarylC₁₋₆alkyloxycarbonyl,        piperazinylcarbonyl, pyrrolidinyl, piperidinylcarbonyl,        C₁₋₆alkyloxycarbonyl, C₁₋₆alkyl substituted with a substituent        selected from hydroxy, amino, aryl, and heteroaryl;        C₃₋₇cycloalkyl substituted with a substituent selected from        hydroxy, amino, aryl, and heteroaryl; piperazinylcarbonyl        substituted with hydroxy, hydroxyC₁₋₆alkyl,        hydroxyC₁₋₆alkyloxyC₁₋₆alkyl; pyrrolidinyl substituted with        hydroxyC₁₋₆alkyl; or piperidinylcarbonyl substituted with one or        two substituents selected from hydroxy, C₁₋₆alkyl,        hydroxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,        C₁₋₆alkyl(dihydroxy)C₁₋₆alkyl or C₁₋₆alkyloxy(hydroxy)C₁₋₆alkyl;    -   R¹⁷ and R¹⁸ are each independently selected from hydrogen,        C₁₋₆alkyl, di(C₁₋₆alkyl)aminoC₁₋₆alkyl, arylC₁₋₆alkyl,        C₁₋₆alkyloxyC₁₋₆alkyl, hydroxyC₁₋₆alkyl,        hydroxyC₁₋₆alkyl(C₁₋₆alkyl) or hydroxyC₁₋₆alkyl(arylC₁₋₆alkyl);

-   -    is —CR⁹═C< and then the dotted line is a bond, —C(═O)—CH<,        —C(═O)—N<, —CHR⁹—CH< or —CHR⁹—N<; wherein    -   each R⁹ is independently hydrogen or C₁₋₆alkyl;

-   R¹ is hydrogen, aryl, heteroaryl, C₁₋₆alkyloxycarbonyl, C₁₋₁₂alkyl,    or C₁₋₁₂alkyl substituted with one or two substituents independently    selected from hydroxy, aryl, heteroaryl, amino, C₁₋₆alkyloxy, mono-    or di(C₁₋₆alkyl)amino, morpholinyl, piperidinyl, pyrrolidinyl,    piperazinyl, C₁₋₆alkylpiperazinyl, arylC₁₋₆alkylpiperazinyl,    heteroarylC₁₋₆alkylpiperazinyl, C₃₋₇cycloalkylpiperazinyl and    C₃₋₇cycloalkylC₁₋₆alkylpiperazinyl;

-   R² is hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy,    heteroarylC₁₋₆alkyloxy, phenylthio, hydroxyC₁₋₆alkylcarbonyl,    C₁₋₆alkyl substituted with a substituent selected from amino, aryl    and heteroaryl; or C₃₋₇cycloalkyl substituted with a substituent    selected from amino, aryl and heteroaryl;

-   R³ is hydrogen, C₁₋₆alkyl, heteroaryl, C₃₋₇cycloalkyl, C₁₋₆alkyl    substituted with a substituent selected from hydroxy, amino, aryl    and heteroaryl; or C₃₋₇cycloalkyl substituted with a substituent    selected from hydroxy, amino, aryl and heteroaryl;

-   R⁴ and R⁵ are each independently hydrogen, halo, C₁₋₆alkyl,    polyhaloC₁₋₆alkyl, cyano, cyanoC₁₋₆alkyl, hydroxy, amino or    C₁₋₆alkyloxy; or

-   R⁴ and R⁵ together can optionally form a bivalent radical selected    from methylenedioxy or ethylenedioxy;

-   R⁶ is hydrogen, C₁₋₆alkyloxycarbonyl or C₁₋₆alkyl;

-   when p is 1 then R⁷ is hydrogen, arylC₁₋₆alkyl, hydroxy or    heteroarylC₁₋₆alkyl;

-   Z is a radical selected from

-   -   wherein    -   each R¹⁰ or R¹¹ are each independently selected from hydrogen,        halo, hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl,        cyano, cyanoC₁₋₆alkyl, tetrazoloC₁₋₆alkyl, aryl, heteroaryl,        arylC₁₋₆alkyl, heteroarylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl,        heteroaryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, heteroarylcarbonyl,        C₁₋₆alkylcarbonyl, arylC₁₋₆alkylcarbonyl,        heteroarylC₁₋₆alkylcarbonyl, C₁₋₆alkyloxy,        C₃₋₇cycloalkylcarbonyl, C₃₋₇cycloalkyl(hydroxy)C₁₋₆alkyl,        arylC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl,        C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,        C₁₋₆alkyloxycarbonylC₁₋₆alkyloxyC₁₋₆alkyl,        hydroxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonylC₂₋₆alkenyl        C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonyl,        C₁₋₆alkylcarbonyloxy, aminocarbonyl, hydroxyC₁₋₆alkyl,        aminoC₁₋₆alkyl, hydroxycarbonyl, hydroxycarbonylC₁₋₆alkyl and        —(CH₂)_(v)—(C(═O)_(r))—(CHR¹⁹)_(u)—NR¹³R¹⁴; wherein        -   v is 0, 1, 2, 3, 4, 5, or 6 and when v is 0 then a direct            bond is intended;        -   r is 0, or 1 and when r is 0 then a direct bond is intended;        -   u is 0, 1, 2, 3, 4, 5, or 6 and when u is 0 then a direct            bond is intended;        -   R¹⁹ is hydrogen or C₁₋₆alkyl;    -   R¹² is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyl        substituted with a substituent selected from hydroxy, amino,        C₁₋₆alkyloxy and aryl; or C₃₋₇cycloalkyl substituted with a        substituent selected from hydroxy, amino, aryl and C₁₋₆alkyloxy;        -   R¹³ and R¹⁴ are each independently selected from hydrogen,            C₁₋₁₂alkyl, C₁₋₆alkylcarbonyl, C₁₋₆alkylsulfonyl,            arylC₁₋₆alkylcarbonyl, C₃₋₇cycloalkyl,            C₃₋₇cycloalkylcarbonyl, —(CH₂)_(k)—NR¹⁵R¹⁶, C₁₋₁₂alkyl            substituted with a substituent selected from hydroxy,            hydroxycarbonyl, cyano, C₁₋₆alkyloxycarbonyl, C₁₋₆alkyloxy,            aryl or heteroaryl; or C₃₋₇cycloalkyl substituted with a            substituent selected from hydroxy, C₁₋₆alkyloxy, aryl,            amino, arylC₁₋₆alkyl, heteroaryl or heteroarylC₁₋₆alkyl; or        -   R¹³ and R¹⁴ together with the nitrogen to which they are            attached can optionally form a morpholinyl, piperidinyl,            pyrrolidinyl, piperazinyl, or piperazinyl substituted with a            substituent selected from C₁₋₆alkyl, arylC₁₋₆alkyl,            arylC₁₋₆alkyloxycarbonyl, heteroarylC₁₋₆alkyl,            C₃₋₇cycloalkyl and C₃₋₇cycloalkylC₁₋₆alkyl; wherein            -   k is 0, 1, 2, 3, 4, 5, or 6 and when k is 0 then a                direct bond is intended;                -   R¹⁵ and R¹⁶ are each independently selected from                    hydrogen, C₁₋₆alkyl, arylC₁₋₆alkyloxycarbonyl,                    C₃₋₇cycloalkyl, C₁₋₁₂alkyl substituted with a                    substituent selected from hydroxy, C₁₋₆alkyloxy,                    aryl, and heteroaryl; and C₃₋₇cycloalkyl substituted                    with a substituent selected from hydroxy,                    C₁₋₆alkyloxy, aryl, arylC₁₋₆alkyl, heteroaryl, and                    heteroarylC₁₋₆alkyl; or                -   R¹⁵ and R¹⁶ together with the nitrogen to which they                    are attached can optionally form a morpholinyl, a                    piperazinyl or a piperazinyl substituted with                    C₁₋₆alkyloxycarbonyl;

-   aryl is phenyl or naphthalenyl;

-   each phenyl or naphthalenyl can optionally be substituted with one,    two or three substituents each independently selected from halo,    hydroxy, C₁₋₆alkyl, amino, polyhaloC₁₋₆alkyl and C₁₋₆alkyloxy; and

-   each phenyl or naphthalenyl can optionally be substituted with a    bivalent radical selected from methylenedioxy and ethylenedioxy;

-   heteroaryl is pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl,    thienyl, oxadiazolyl, tetrazolyl, benzofuranyl or tetrahydrofuranyl;

-   each pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,    oxadiazolyl, tetrazolyl, benzofuranyl, or tetrahydrofuranyl can    optionally be substituted with one, two or three substituents each    independently selected from halo, hydroxy, C₁₋₆alkyl, amino,    polyhaloC₁₋₆alkyl, aryl, arylC₁₋₆alkyl or C₁₋₆alkyloxy; and

-   each pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,    benzofuranyl, or tetrahydrofuranyl can optionally be substituted    with a bivalent radical selected from methylenedioxy or    ethylenedioxy;

-   with the proviso that

-   when m is 1; the substituents on the phenyl ring other than R² are    in the meta position;

-   s is 0; and t is 0; then

-   Z is a radical selected from (a-1), (a-3), (a-4), (a-5), (a-6),    (a-7), (a-8) or (a-9).

The compounds of formula (I) may also exist in their tautomeric forms.Such forms although not explicitly indicated in the above formula areintended to be included within the scope of the present invention.

A number of terms used in the foregoing definitions and hereinafter areexplained hereunder. These terms are sometimes used as such or incomposite terms.

As used in the foregoing definitions and hereinafter, halo is generic tofluoro, chloro, bromo and iodo; C₁₋₆alkyl defines straight and branchedchain saturated hydrocarbon radicals having from 1 to 6 carbon atomssuch as, e.g. methyl, ethyl, propyl, butyl, pentyl, hexyl,1-methylethyl, 2-methylpropyl, 2-methyl-butyl, 2-methylpentyl and thelike; C₁₋₆alkanediyl defines bivalent straight and branched chainedsaturated hydrocarbon radicals having from 1 to 6 carbon atoms such as,for example, methylene, 1,2-ethanediyl, 1,3-propanediyl 1,4-butanediyl,1,5-pentanediyl, 1,6-hexanediyl and the branched isomers thereof suchas, 2-methylpentanediyl, 3-methylpentanediyl, 2,2-dimethylbutanediyl,2,3-dimethylbutanediyl and the like; C₁₋₁₂ alkyl includes C₁₋₆alkyl andthe higher homologues thereof having 7 to 12 carbon atoms such as, forexample heptyl, octyl, nonyl, decyl, undecyl and dodecyl;hydroxyC₁₋₆alkyl defines a hydroxy substituent on straight and branchedchain saturated hydrocarbon radicals having from 1 to 6 carbon atoms;trihalomethyl defines methyl containing three identical or differenthalo substituents for example trifluoromethyl; C₂₋₆alkenyl definesstraight and branched chain hydrocarbon radicals containing one doublebond and having from 2 to 6 carbon atoms such as, for example, ethenyl,2-propenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 3-methyl-2-butenyl, andthe like; C₃₋₇alkynyl defines straight and branched chained hydrocarbonradicals containing one triple bond and having from 3 to 6 carbon atoms,such as, for example, 2-propynyl, 3-butynyl, 2-butynyl, 2-pentynyl,3-pentynyl, 3-hexynyl, and the like; C₃₋₇cycloalkyl includes cyclichydrocarbon groups having from 3 to 10 carbons, such as cyclopropyl,cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl,cycloheptyl, and the like.

The term “addition salt” comprises the salts which the compounds offormula (I) are able to form with organic or inorganic bases such asamines, alkali metal bases and earth alkaline metal bases, or quaternaryammonium bases, or with organic or inorganic acids, such as mineralacids, sulfonic acids, carboxylic acids or phosphorus containing acids.

The term “addition salt” further comprises pharmaceutically acceptablesalts, metal complexes and solvates and the salts thereof, that thecompounds of formula (I) are able to form.

The term “pharmaceutically acceptable salts” means pharmaceuticallyacceptable acid or base addition salts. The pharmaceutically acceptableacid or base addition salts as mentioned hereinabove are meant tocomprise the therapeutically active non-toxic acid and non-toxic baseaddition salt forms which the compounds of formula (I) are able to form.The compounds of formula (I) which have basic properties can beconverted in their pharmaceutically acceptable acid addition salts bytreating said base form with an appropriate acid. Appropriate acidscomprise, for example, inorganic acids such as hydrohalic acids, e.g.hydrochloric or hydrobromic acid; sulfuric; nitric; phosphoric and thelike acids; or organic acids such as, for example, acetic, propanoic,hydroxyacetic, lactic, pyruvic, oxalic, malonic, succinic (i.e.butanedioic acid), maleic, fumaric, malic, tartaric, citric,methanesulfonic, ethanesulfonic, benzenesulfonic, p-toluenesulfonic,cyclamic, salicylic, p-aminosalicylic, pamoic and the like acids.

The compounds of formula (I) which have acidic properties may beconverted in their pharmaceutically acceptable base addition salts bytreating said acid form with a suitable organic or inorganic base.Appropriate base salt forms comprise, for example, the ammonium salts,the alkali and earth alkaline metal salts, e.g. the lithium, sodium,potassium, magnesium, calcium salts and the like, salts with organicbases, e.g. the benzathine, N-methyl-D-glucamine, hydrabamine salts, andsalts with amino acids such as, for example, arginine, lysine and thelike.

The terms acid or base addition salt also comprise the hydrates and thesolvent addition forms which the compounds of formula (I) are able toform. Examples of such forms are e.g. hydrates, alcoholates and thelike.

The term “metal complexes” means a complex formed between a compound offormula (I) and one or more organic or inorganic metal salt or salts.Examples of said organic or inorganic salts comprise the halogenides,nitrates, sulfates, phosphates, acetates, trifluoroacetates,trichloroacetates, propionates, tartrates, sulfonates, e.g.methylsulfonates, 4-methylphenylsulfonates, salicylates, benzoates andthe like of the metals of the second main group of the periodicalsystem, e.g. the magnesium or calcium salts, of the third or fourth maingroup, e.g. aluminium, tin, lead as well as the first to the eighthtransition groups of the periodical system such as, for example,chromium, manganese, iron, cobalt, nickel, copper, zinc and the like.

The term “stereochemically isomeric forms of compounds of formula (I)”,as used hereinbefore, defines all possible compounds made up of the sameatoms bonded by the same sequence of bonds but having differentthree-dimensional structures which are not interchangeable, which thecompounds of formula (I) may possess. Unless otherwise mentioned orindicated, the chemical designation of a compound encompasses themixture of all possible stereochemically isomeric forms which saidcompound may possess. Said mixture may contain all diastereomers and/orenantiomers of the basic molecular structure of said compound. Allstereochemically isomeric forms of the compounds of formula (I) both inpure form or in admixture with each other are intended to be embracedwithin the scope of the present invention.

The N-oxide forms of the compounds of formula (I) are meant to comprisethose compounds of formula (I) wherein one or several nitrogen atoms areoxidized to the so-called N-oxide, particularly those N-oxides whereinone or more of the piperidine piperazine or pyridazinyl-nitrogens areN-oxidized.

Whenever used hereinafter, the term “compounds of formula (I)” is meantto include also the N-oxide forms, the pharmaceutically acceptable acidor base addition salts and all stereoisomeric forms.

A first group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   a) X is C(═O) or CHR⁸; wherein    -   R⁸ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, aminocarbonyl, mono-        or di(C₁₋₆alkyl)aminocarbonyl, hydroxycarbonyl,        arylC₁₋₆alkyloxycarbonyl, heteroarylC₁₋₆alkyloxycarbonyl,        C₁₋₆alkyloxycarbonyl, C₁₋₆alkyl substituted with a substituent        selected from hydroxy, amino, aryl, and heteroaryl or        C₃₋₇cycloalkyl substituted with a substituent selected from        hydroxy, amino, aryl and heteroaryl;-   b) R¹ is hydrogen, aryl, heteroaryl, C₁₋₁₂alkyl, or C₁₋₁₂alkyl    substituted with one or two substituents independently selected from    hydroxy, aryl, heteroaryl, amino, C₁₋₆alkyloxy, mono- or    di(C₁₋₆alkyl)amino, morpholinyl, piperidinyl, pyrrolidinyl,    piperazinyl, C₁₋₆alkylpiperazinyl, arylC₁₋₆alkylpiperazinyl,    heteroarylC₁₋₆alkylpiperazinyl, C₃₋₇cycloalkylpiperazinyl and    C₃₋₇cycloalkylC₁₋₆alkylpiperazinyl;-   c) R³ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyl substituted    with a substituent selected from hydroxy, amino, aryl, and    heteroaryl; or C₃₋₇cycloalkyl substituted with a substituent    selected from hydroxy, amino, aryl and heteroaryl;-   d) R⁴ and R⁵ are each independently hydrogen, halo, C₁₋₆alkyl,    polyhaloC₁₋₆alkyl, hydroxy, amino or C₁₋₆alkyloxy;-   e) R⁴ and R⁵ together can optionally form a bivalent radical    selected from methylenedioxy or ethylenedioxy;-   f) R⁶ is hydrogen or C₁₋₆alkyl;-   g) when p is 1 then R⁷ is hydrogen, arylC₁₋₆alkyl or    heteroarylC₁₋₆alkyl;-   h) Z is a radical selected from (a-1), (a-2), (a-3), (a-4), (a-5)    and (a-6);-   i) each R¹⁰ or R¹¹ are each independently selected from hydrogen,    hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano,    cyanoC₁₋₆alkyl, tetrazoloC₁₋₆alkyl, aryl, heteroaryl, arylC₁₋₆alkyl,    heteroarylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl,    heteroaryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, heteroarylcarbonyl,    arylC₁₋₆alkylcarbonyl, heteroarylC₁₋₆alkylcarbonyl, C₁₋₆alkyloxy,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonyl, C₁₋₆alkylcarbonyloxy,    aminocarbonyl, hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl,    hydroxycarbonylC₁₋₆alkyl and    —(CH₂)_(v)—(C(═O)_(r))—(CH₂)_(u)—NR¹³R¹⁴;-   j) R¹³ and R¹⁴ are each independently selected from hydrogen,    C₁₋₁₂alkyl, C₃₋₇cycloalkyl, —(CH₂)_(k)—NR¹⁵R¹⁶, C₁₋₁₂alkyl    substituted with a substituent selected from hydroxy, C₁₋₆alkyloxy,    aryl, and heteroaryl; or C₃₋₇cycloalkyl substituted with a    substituent selected from hydroxy, C₁₋₆alkyloxy, aryl,    arylC₁₋₆alkyl, heteroaryl and heteroarylC₁₋₆alkyl;-   k) R¹³ and R¹⁴ together with the nitrogen to which they are attached    can optionally form a morpholinyl, piperidinyl, pyrrolidinyl,    piperazinyl, or piperazinyl substituted with a substituent selected    from C₁₋₆alkyl, arylC₁₋₆alkyl, heteroarylC₁₋₆alkyl, C₃₋₇cycloalkyl,    and C₃₋₇cycloalkylC₁₋₆alkyl;-   l) R¹⁵ and R¹⁶ are each independently selected from hydrogen,    C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₁₂alkyl substituted with a substituent    selected from hydroxy, C₁₋₆alkyloxy, aryl, and heteroaryl; and    C₃₋₇cycloalkyl substituted with a substituent selected from hydroxy,    C₁₋₆alkyloxy, aryl, arylC₁₋₆alkyl, heteroaryl and    heteroarylC₁₋₆alkyl;-   m) heteroaryl is pyridinyl, indolyl, quinolinyl, imidazolyl,    furanyl, thienyl, benzofuranyl, or tetrahydrofuranyl; and each    pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,    benzofuranyl, or tetrahydrofuranyl can optionally be substituted    with one, two or three substituents each independently selected from    halo, hydroxy, C₁₋₆alkyl, amino, polyhaloC₁₋₆alkyl and C₁₋₆alkyloxy;    and-   n) each pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl,    thienyl, benzofuranyl, or tetrahydrofuranyl can optionally be    substituted with a bivalent radical selected from methylenedioxy or    ethylenedioxy;

A second group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   a) n is 0, 1 or 2;-   b) p is 0;-   c) R⁸ is hydrogen, aminocarbonyl, arylC₁₋₆alkyloxycarbonyl or    C₁₋₆alkyl substituted with hydroxy;-   d)

-    is —CR⁹═C< or —CHR⁹—CH<;-   e) R¹ is hydrogen, C₁₋₁₂alkyl, or C₁₋₁₂alkyl substituted with    heteroaryl;-   f) R² is hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy    or phenylthio;-   g) R³ is hydrogen or C₁₋₆alkyl;-   h) R⁴ and R⁵ are each independently hydrogen, halo or C₁₋₆alkyloxy;-   i) Z is a radical selected from (a-1), (a-2), (a-3), (a-4) or (a-6);-   j) each R¹⁰ or R¹¹ are independently selected from hydrogen,    hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano, aryl,    arylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, C₁₋₆alkyloxy,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonyl, aminocarbonyl,    hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl and    —(CH₂)_(v)—(C(═O)_(r))—(CH₂)_(u)—NR¹³R¹⁴;-   k) v is 0, or 1;-   l) r is o or 1;-   m) u is 0;-   n) R¹³ and R¹⁴ are each independently selected from hydrogen,    C₁₋₆alkyl, —(CH₂)_(k)—NR¹⁵R¹⁶ and C₁₋₁₂alkyl substituted with    hydroxy;-   o) R¹³ and R¹⁴ together with the nitrogen to which they are attached    can form a pyrrolidinyl;-   p) k is 2;-   q) R¹⁵ and R¹⁶ are each independently C₁₋₆alkyl;-   r) aryl is phenyl or phenyl substituted with halo; and-   s) heteroaryl is pyridinyl or indolyl.

A third group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   a) m is 0 or 2;-   b) n is 0, 2 or 3;-   c) p is 1;-   d) s is 1;-   e) t is 1;-   f) X is C(═O);-   g)

-    is —C(═O)—CH<, —C(═O)—N<, —CHR⁹—CH<, or —CHR⁹—N<;-   h) R¹ is aryl, heteroaryl, C₁₋₆alkyloxycarbonyl, C₁₋₁₂alkyl, or    C₁₋₁₂alkyl substituted with one or two substituents independently    selected from hydroxy, aryl, heteroaryl, amino, C₁₋₆alkyloxy, mono-    or di(C₁₋₆alkyl)amino, morpholinyl, piperidinyl, pyrrolidinyl,    piperazinyl, C₁₋₆alkylpiperazinyl, aryl C₁₋₆alkylpiperazinyl,    heteroarylC₁₋₆alkylpiperazinyl, C₃₋₇cycloalkylpiperazinyl and    C₃₋₇cycloalkylC₁₋₆alkylpiperazinyl;-   i) R² is halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy,    heteroarylC₁₋₆alkyloxy, phenylthio, hydroxyC₁₋₆alkylcarbonyl,    C₁₋₆alkyl substituted with a substituent selected from amino, aryl    and heteroaryl; or C₁₋₇cycloalkyl substituted with a substituent    selected from amino, aryl and heteroaryl;-   j) R³ is C₁₋₆alkyl, C₃₋₇cycloalkyl, C₁₋₆alkyl substituted with a    substituent selected from hydroxy, amino, aryl, and heteroaryl; or    C₃₋₇cycloalkyl substituted with a substituent selected from hydroxy,    amino, aryl and heteroaryl;-   k) R⁴ and R⁵ are each independently C₁₋₆alkyl, polyhaloC₁₋₆alkyl,    cyano, cyanoC₁₋₆alkyl, hydroxy or amino;-   l) R⁴ and R⁵ together can optionally form a bivalent radical    selected from methylenedioxy or ethylenedioxy;-   m) R⁶ is C₁₋₆alkyloxycarbonyl or C₁₋₆alkyl;-   n) R⁷ is hydrogen, arylC₁₋₆alkyl, hydroxy or heteroarylC₁₋₆alkyl;    and-   o) Z is a radical selected from (a-1), (a-3), (a-4), (a-5), (a-6),    (a-7), (a-8) or (a-9).

A fourth group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   a) R¹⁸ is hydrogen, —C(═O)—NR¹⁷R¹⁸, arylC₁₋₆alkyloxycarbonyl,    C₁₋₆alkyl substituted with hydroxy, piperazinylcarbonyl substituted    with hydroxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl,    pyrrolidinyl substituted with hydroxyC₁₋₆alkyl or    piperidinylcarbonyl substituted with one or two substituents    selected from hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyl(dihydroxy)C₁₋₆alkyl or    C₁₋₆alkyloxy(hydroxy)C₁₋₆alkyl;-   b) R¹⁷ and R¹⁸ are each independently selected from hydrogen,    C₁₋₆alkyl, di(C₁₋₆alkyl)aminoC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl or    hydroxyC₁₋₆alkyl;-   c)

-    is —CR⁹═C< and then the dotted line is a bond, —CHR⁹—CH< or    —CHR⁹—N<;-   d) R¹ is hydrogen, heteroaryl, C₁₋₆alkyloxycarbonyl, C₁₋₁₂alkyl or    C₁₋₁₂alkyl substituted with heteroaryl;-   e) R² is hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy    or phenylthio;-   f) R³ is hydrogen, C₁₋₆alkyl or heteroaryl;-   g) R⁴ and R⁵ are each independently hydrogen, halo, C₁₋₆alkyl,    cyano, cyanoC₁₋₆alkyl, hydroxy or C₁₋₆alkyloxy;-   h) when p is 1 then R⁷ is arylC₁₋₆alkyl or hydroxy;-   i) Z is a radical selected from (a-1), (a-2), (a-3), (a-4), (a-5),    (a-6), (a-8), (a-9), (a-10) and (a-11);-   j) each R¹⁰ or R¹¹ are each independently selected from hydrogen,    halo, hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano,    cyanoC₁₋₆alkyl, tetrazoloC₁₋₆alkyl, aryl, heteroaryl,    heteroarylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl, arylcarbonyl,    C₁₋₆alkylcarbonyl, C₃₋₇ cycloalkylcarbonyl,    C₃₋₇cycloalkyl(hydroxy)C₁₋₆alkyl, arylC₁₋₆alkyloxyC₁₋₆alkyl,    C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,    C₁₋₆alkyloxycarbonylC₁₋₆alkyloxyC₁₋₆alkyl,    hydroxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonylC₂₋₆alkenyl,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonyl, aminocarbonyl,    hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl,    hydroxycarbonylC₁₋₆alkyl and    —(CH₂)_(v)—(C(═O)_(r))—(CHR¹⁹)_(u)—NR¹³R¹⁴;-   k) v is 0 or 1;-   l) u is 0 or 1;-   m) R¹² is hydrogen or C₁₋₆alkyl;-   n) R¹³ and R¹⁴ are each independently selected from hydrogen,    C₁₋₁₂alkyl, C₁₋₆alkylcarbonyl, C₁₋₆alkylsulfonyl,    arylC₁₋₆alkylcarbonyl, C₃₋₇cycloalkylcarbonyl, —(CH₂)_(k)—NR¹⁵R¹⁶,    C₁₋₁₂alkyl substituted with a substituent selected from hydroxy,    hydroxycarbonyl, cyano, C₁₋₆alkyloxycarbonyl or aryl;-   o) R¹³ and R¹⁴ together with the nitrogen to which they are attached    can optionally form a morpholinyl, pyrrolidinyl, piperazinyl, or    piperazinyl substituted with a substituent selected from C₁₋₆alkyl    or arylC₁₋₆alkyloxycarbonyl;-   p) k is 2;-   q) R¹⁵ and R¹⁶ are each independently selected from hydrogen,    C₁₋₆alkyl or arylC₁₋₆alkyloxycarbonyl;-   r) R¹⁵ and R¹⁶ together with the nitrogen to which they are attached    can optionally form a morpholinyl, a piperazinyl or a piperazinyl    substituted with C₁₋₆alkyloxycarbonyl;-   s) aryl is phenyl or phenyl substituted with halo;-   t) heteroaryl is pyridinyl, indolyl, oxadiazolyl or tetrazolyl; and-   u) each pyridinyl, indolyl, oxadiazolyl or tetrazolyl can optionally    be substituted with one substituents selected from C₁₋₆alkyl, aryl    or arylC₁₋₆alkyl.

A fifth group of interesting compounds consists of those compounds offormula (I) wherein one or more of the following restrictions apply:

-   a) m is 0;-   b) n is 1;-   c) p is 0;-   d) s is 0;-   e) t is 0;-   f) X is CHR⁸;-   g) R⁸ is hydrogen;-   h)

-    is —CR⁹═C<;-   i) each R⁹ is hydrogen;-   j) R¹ is hydrogen;-   k) R² is hydrogen or C₁₋₆alkyloxy;-   l) R³ is hydrogen;-   m) R⁴ and R⁵ are each independently hydrogen, C₁₋₆alkyl or    C₁₋₆alkyloxy;-   n) R⁶ is hydrogen;-   o) Z is a radical selected from (a-1), (a-2), (a-3) or (a-4);-   p) R¹⁰ or R¹¹ are each independently selected from hydrogen, hydroxy    or hydroxyC₁₋₆alkyl.

A group of preferred compounds consists of those compounds of formula(I) or any subgroup thereof, wherein

-   R⁸ is hydrogen, —C(═O)—NR¹⁷R¹⁸, arylC₁₋₆alkyloxycarbonyl, C₁₋₆alkyl    substituted with hydroxy, piperazinylcarbonyl substituted with    hydroxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl,    pyrrolidinyl substituted with hydroxyC₁₋₆alkyl or    piperidinylcarbonyl substituted with one or two substituents    selected from hydroxy, C₁₋₆alkyl, hydroxyC₁₋₆alkyl,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyl(dihydroxy)C₁₋₆alkyl or    C₁₋₆alkyloxy(hydroxy)C₁₋₆alkyl; R¹⁷ and R¹⁸ are each independently    selected from hydrogen, C₁₋₆alkyl, di(C₁₋₆alkyl)aminoC₁₋₆alkyl,    arylC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl or hydroxyC₁₋₆alkyl;

-    is —CR⁹═C< and then the dotted line is a bond, —CHR⁹—CH< or    —CHR⁹—N<; R¹ is hydrogen, heteroaryl, C₁₋₆alkyloxycarbonyl,    C₁₋₁₂alkyl or C₁₋₁₂alkyl substituted with heteroaryl; R² is    hydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy or    phenylthio; R³ is hydrogen, C₁₋₆alkyl or heteroaryl; R⁴ and R⁵ are    each independently hydrogen, halo, C₁₋₆alkyl, cyano, cyanoC₁₋₆alkyl,    hydroxy or C₁₋₆alkyloxy; when p is 1 then R⁷ is arylC₁₋₆alkyl or    hydroxy; Z is a radical selected from (a-1), (a-2), (a-3), (a-4),    (a-5), (a-6), (a-8), (a-9), (a-10) and (a-11); each R¹⁰ or R¹¹ are    each independently selected from hydrogen, halo, hydroxy, amino,    C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano, cyanoC₁₋₆alkyl,    tetrazoloC₁₋₆alkyl, aryl, heteroaryl, heteroarylC₁₋₆alkyl,    aryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, C₁₋₆alkylcarbonyl,    C₃₋₇cycloalkylcarbonyl, C₃₋₇cycloalkyl(hydroxy)C₁₋₆alkyl,    arylC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl,    C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,    C₁₋₆alkyloxycarbonylC₁₋₆alkyloxyC₁₋₆alkyl,    hydroxyC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonylC₂₋₆alkenyl,    C₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxycarbonyl, aminocarbonyl,    hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl,    hydroxycarbonylC₁₋₆alkyl and    —(CH₂)_(v)—(C(═O)_(r))—(CHR¹⁹)_(u)—NR¹³R¹⁴; v is 0 or 1; u is 0 or    1; R¹² is hydrogen or C₁₋₆alkyl; R¹³ and R¹⁴ are each independently    selected from hydrogen, C₁₋₁₂alkyl, C₁₋₆alkylcarbonyl,    C₁₋₆alkylsulfonyl, arylC₁₋₆alkylcarbonyl, C₃₋₇cycloalkylcarbonyl,    —(CH₂)_(k)—NR¹⁵R¹⁶, C₁₋₁₂alkyl substituted with a substituent    selected from hydroxy, hydroxycarbonyl, cyano, C₁₋₆alkyloxycarbonyl    or aryl; R¹³ and R¹⁴ together with the nitrogen to which they are    attached can optionally form a morpholinyl, pyrrolidinyl,    piperazinyl or piperazinyl substituted with a substituent selected    from C₁₋₆alkyl or arylC₁₋₆alkyloxycarbonyl; k is 2; R¹⁵ and R¹⁶ are    each independently selected from hydrogen, C₁₋₆alkyl or    arylC₁₋₆alkyloxycarbonyl; k is 2; R¹⁵ and R¹⁶ are each independently    selected from hydrogen, C₁₋₆alkyl or arylC₁₋₆alkyloxycarbonyl; R¹⁵    and R¹⁶ together with the nitrogen to which they are attached can    optionally form a morpholinyl or piperazinyl, or piperazinyl    substituted with C₁₋₆alkyloxycarbonyl; aryl is phenyl or phenyl    substituted with halo; heteroaryl is pyridinyl, indolyl, oxadiazolyl    or tetrazolyl; and each pyridinyl, indolyl, oxadiazolyl or    tetrazolyl can optionally be substituted with one substituent    selected from C₁₋₆alkyl, aryl or arylC₁₋₆alkyl.

A group of more preferred compounds consists of those compounds offormula (I) or any subgroup thereof wherein m is 0; n is 1; p is 0; s is0; t is 0; X is CHR⁸; R⁸ is hydrogen;

is —CR⁹═C<; each R⁹ is hydrogen; R¹ is hydrogen; R² is hydrogen orC₁₋₆alkyloxy; R² is hydrogen or C₁₋₆alkyloxy; R³ is hydrogen; R⁴ and R⁵are each independently hydrogen, C₁₋₆alkyl or C₁₋₆alkyloxy; R⁶ ishydrogen; Z is a radical selected from (a-1), (a-2), (a-3) or (a-4); andR¹⁰ or R¹¹ are each independently selected from hydrogen, hydroxy orhydroxyC₁₋₆alkyl.

The most preferred compounds are compound No. 1, compound No. 21,compound No. 4, compound No. 5, compound No. 36, compound No. 69,compound No. 110, compound No. 111, compound No. 112, compound No. 229and compound No. 37.

The compounds of formula (I), their pharmaceutically acceptable saltsand N-oxides and stereochemically isomeric forms thereof may be preparedin conventional manner. The starting materials and some of theintermediates are known compounds and are commercially available or maybe prepared according to conventional reaction procedures as generallyknown in the art.

A number of such preparation methods will be described hereinafter inmore detail. Other methods for obtaining final compounds of formula (I)are described in the examples.

The compounds of formula (I) can be prepared by reacting an intermediateof formula (II) with an intermediate of formula (III) wherein W is anappropriate leaving group such as, for example, halo, e.g. fluoro,chloro, bromo or iodo, or a sulfonyloxy radical such asmethylsulfonyloxy, 4-methylphenylsulfonyloxy and the like. The reactioncan be performed in a reaction-inert solvent such as, for example, analcohol, e.g. methanol, ethanol, 2-methoxy-ethanol, propanol, butanoland the like; an ether, e.g. 4,4-dioxane, 1,1′-oxybispropane and thelike; a ketone, e.g. 4-methyl-2-pentanone; or N,N-dimethylformamide,nitrobenzene, acetonitrile, acetic acid and the like. The addition of anappropriate base such as, for example, an alkali or earth alkaline metalcarbonate or hydrogen carbonate, e.g. triethylamine or sodium carbonate,may be utilized to pick up the acid which is liberated during the courseof the reaction. A small amount of an appropriate metal iodide, e.g.,sodium or potassium iodide may be added to promote the reaction.Stirring may enhance the rate of the reaction. The reaction mayconveniently be carried out at a temperature ranging between roomtemperature and the reflux temperature of the reaction mixture and, ifdesired, the reaction may be carried out at an increased pressure.

The compounds of formula (I), wherein X is CH₂, herein referred to ascompounds of formula (I-a), can be prepared by converting compounds offormula (I) wherein X is C(═O), herein referred to as compounds offormula (I-b), by reacting the compound of formula (I-b) with lithiumaluminium hydride in a suitable solvent such as tetrahydrofuran.

The compounds of formula (I-a) can also be prepared by reacting anappropriate carboxaldehyde of formula (IV), with an intermediate offormula (V), in the presence of an appropriate reagent, such as a sodiumborohydride e.g. sodium tetrahydroborate or polymer supportedcyanotrihydroborate, in a suitable solvent, such as an alcohol e.g.methanol.

In an identical way the compounds of formula (I), wherein t is 1, hereinreferred to as compounds of formula (I-c), can be prepared by reactingan intermediate of formula (II) with an appropriate carboxaldehyde offormula (VI).

The compounds of formula (I), wherein s is 1, herein referred to ascompounds of formula (I-d), can be prepared by reacting an intermediateof formula (VII) with lithium aluminium hydride in a suitable solventsuch as tetrahydrofuran.

The compounds of formula (I) and the intermediates of formula (III) mayalso be converted into each other via art-known reactions or functionalgroup transformations. A number of such transformations are alreadydescribed hereinabove. Other examples are hydrolysis of carboxylicesters to the corresponding carboxylic acid or alcohol; hydrolysis ofamides to the corresponding carboxylic acids or amines; hydrolysis ofnitriles to the corresponding amides; amino groups on imidazole orphenyl may be replaced by a hydrogen by art-known diazotation reactionsand subsequent replacement of the diazo-group by hydrogen; alcohols maybe converted into esters and ethers; primary amines may be convertedinto secondary or tertiary amines; double bonds may be hydrogenated tothe corresponding single bond; an iodo radical on a phenyl group may beconverted in to an ester group by carbon monoxide insertion in thepresence of a suitable palladium catalyst.

Intermediates of formula (II), wherein X is CH₂, m is 0, s is 0 and R³is hydrogen, herein referred to as intermediates of formula (II-a), canbe prepared by a nitro to amine reduction reaction starting with anintermediate of formula (VIII), in the presence of a metal catalyst suchas Raney Nickel and an appropriate reductant such as hydrogen, in asuitable solvent such as methanol or ethanol.

Intermediates of formula (II), wherein X is C(═O), s is 0 and R³ ishydrogen, herein referred to as intermediates of formula (II-b), can beprepared by reacting an intermediate of formula (IX) with anintermediate of formula (X) in the presence of appropriate reagents suchas N′-(ethylcarbonimidoyl)-N,N-dimethyl-1,3-propanediamine,monohydrochloride (EDC) and 1-hydroxy-1H-benzotriazole (HOBT). Thereaction may be performed in the presence of a base such astriethylamine, in a suitable solvent, such as, a mixture ofdichloromethane and tetrahydrofuran.

The intermediates of formula (IV) can be prepared by reactingintermediates of formula (XI) with lithium aluminium hydride in asuitable solvent such as tetrahydrofuran.

The intermediates of formula (VII) can be prepared by reacting anintermediate of formula (XII) with an intermediate of formula (XIII) inthe presence of 2-Chloro-1-methylpyridinium iodide and triethylamine ina suitable solvent such as acetonitrile.

The intermediates of formula (VIII) can be prepared by reacting anintermediate of formula (XIV) with an intermediate of formula (XV),wherein A is an appropriate leaving group such as, for example, halo,e.g. fluoro, chloro, bromo or iodo, or C₁₋₆alkyloxy, e.g. methyloxy, indiisopropylethyl amine.

The intermediates of formula (XII) can be prepared by converting anintermediate of formula (XVI) in the presence of sodium hydroxide andwater, in a suitable solvent, such as ethanol.

The intermediates of formula (XVI) can be prepared by reacting anintermediate of formula (XVIII), wherein A is as defined above, with anintermediate of formula (XIV), in a suitable solvent such asdiisopropylethyl amine.

The compounds of formula (I) and some of the intermediates may have atleast one stereogenic centre in their structure. Such stereogenic centremay be present in an R or an S configuration.

The compounds of formula (I) as prepared in the hereinabove describedprocesses are generally racemic mixtures of enantiomers, which can beseparated from one another following art-known resolution procedures.The racemic compounds of formula (I) may be converted into thecorresponding diastereomeric salt forms by reaction with a suitablechiral acid. Said diastereomeric salt forms are subsequently separated,for example, by selective or fractional crystallization and theenantiomers are liberated there from by alkali. An alternative manner ofseparating the enantiomeric forms of the compounds of formula (I)involves liquid chromatography using a chiral stationary phase. Saidpure stereochemically isomeric forms may also be derived from thecorresponding pure stereochemically isomeric forms of the appropriatestarting materials, provided that the reaction occursstereospecifically. Preferably if a specific stereoisomer is desired,said compound would be synthesized by stereospecific methods ofpreparation. These methods will advantageously employ enantiomericallypure starting materials.

The compounds of formula (I), the pharmaceutically acceptable acidaddition salts and stereoisomeric forms thereof have valuablepharmacological properties in that they inhibit the interaction betweenp53 and MDM2.

The term “MDM2” is used herein to mean a protein obtained as a result ofexpression of the mdm2 gene. Within the meaning of this term, MDM2encompass all proteins encoded by mdm2, mutants thereof, alternativeslice proteins thereof, and phosphorylated proteins thereof.Additionally, as used herein, the term “MDM2” includes MDM2 analogues,e.g. MDMX, also known as MDM4, and MDM2 homologues and analogues ofother animals, e.g. the human homologue HDM2 or the human analogue HDMX.

The term “inhibiting the interaction” or “inhibitor of the interaction”is used herein to mean preventing or reducing the direct of indirectassociation of one or more molecules, peptides, proteins, enzymes orreceptors; or preventing or reducing the normal activity of one or moremolecules, peptides, proteins, enzymes, or receptors.

The term “inhibitor of the interaction of p53 with MDM2” or “p53-MDM2inhibitor” is used herein to describe an agent which increases theexpression of p53 in the assay described in C.1. This increase may becaused by, but is not limited to, one or more of the followingmechanisms of action:

-   -   inhibiting the interaction between p53 and MDM2,    -   direct association with either the MDM2 or the p53 protein,    -   interactions with upstream or downstream targets, e.g. kinases,        or enzyme activities involved in ubiquitination or SUMO        modification,    -   sequestering or transportation of MDM2 and p53 into different        cellular compartments,    -   modulation of proteins associating with MDM2, for example (but        not limited to), p73, E2F-1, Rb, p21 waf1 or cip1,    -   downregulating or interference with MDM2 expression and/or MDM2        activity, for example by (but not limited to), impacting on its        cellular localisation, post-translational modification, nuclear        export or ubiquitin ligase activity    -   direct or indirect stabilization of the p53 protein, e.g. by        keeping it in its functional structural form, or by preventing        misfolding,    -   enhancing p53 expression or expression of p53 family members,        e.g. p63 and p73.    -   increasing p53 activity, for example by (but not limited to),        enhancing its transcriptional activity and/or    -   increasing expression of genes and proteins of the        p53-signalling pathway, for example (but not limited to) p21        waf1, cip1, MIC-1 (GDF-15), PIG-3 and ATF-3.

Hence, the present invention discloses the compounds of formula (I) foruse as a medicine.

Furthermore, the invention also concerns the use of a compound for themanufacture of a medicament for the treatment of a disorder mediatedthrough a p53-MDM2 interaction, wherein said compound is a compound offormula (I)

The term “treating” or “treatment” as used herein covers any treatmentof a disease and/or condition in an animal, particularly a human, andincludes: (i) preventing a disease and/or condition from occurring in asubject which may be predisposed to the disease and/or condition but hasnot yet been diagnosed as having it; (ii) inhibiting the disease and/orcondition, i.e., arresting its development; (iii) relieving the diseaseand/or condition, i.e., causing regression of the disease and/orcondition.

With the term “a disorder mediated through a p53-MDM2 interaction” ismeant any undesired or detrimental condition that results in or from theinhibition of the interaction between the MDM2 protein and p53 or othercellular proteins that induce apoptosis, induce cellular death, orregulate the cell cycle.

This invention also provides a method for treating a disorder mediatedthrough a p53-MDM2 interaction by administering an effective amount of acompound of the present invention, to a subject, e.g. a mammal (and moreparticularly a human) in need of such treatment.

The compounds of the invention can have antiproliferative effects intumour cells, even if such cells are devoid of functional p53. More inparticular, the compounds of the invention can have antiproliferativeeffects in tumours with wild-type p53 and/or in tumours overexpressingMDM2.

Thus, this invention also provides a method for inhibiting tumour growthby administering an effective amount of a compound of the presentinvention, to a subject, e.g. a mammal (and more particularly a human)in need of such treatment.

Examples of tumours which may be inhibited, but are not limited to, lungcancer (e.g. adenocarcinoma and including non-small cell lung cancer),pancreatic cancers (e.g. pancreatic carcinoma such as, for exampleexocrine pancreatic carcinoma), colon cancers (e.g. colorectalcarcinomas, such as, for example, colon adenocarcinoma and colonadenoma), oesophageal cancer, oral squamous carcinoma, tongue carcinoma,gastric carcinoma, nasopharyngeal cancer, hematopoietic tumours oflymphoid lineage (e.g. acute lymphocytic leukemia, B-cell lymphoma,Burkitt's lymphoma), myeloid leukemias (for example, acute myelogenousleukemia (AML)), thyroid follicular cancer, myclodysplastic syndrome(MDS), tumours of mesenchymal origin (e.g. fibrosarcomas andrhabdomyosarcomas), melanomas, teratocarcinomas, neuroblastomas, braintumors, gliomas, benign tumour of the skin (e.g. keratoacanthomas),breast carcinoma (e.g. advanced breast cancer), kidney carcinoma, ovarycarcinoma, cervical carcinoma, endometrial carcinoma, bladder carcinoma,prostate cancer including the advanced disease, testicular cancers,osteosarcoma, head and neck cancer and epidermal carcinoma.

The compounds of the present invention can also be used for thetreatment and prevention of inflammatory conditions.

Thus, this invention also provides a method for the treatment andprevention of inflammatory conditions by administering an effectiveamount of a compound of the present invention, to a subject, e.g. amammal (and more particularly a human) in need of such treatment.

The compounds of the present invention can also be used for thetreatment of autoimmune diseases and conditions. With the term“autoimmune diseases” is meant any disease in which an animal's immunesystem reacts adversely to a self-antigen. With the term “self-antigen”is meant any antigen that is normally found in the animal's body.Representative autoimmune diseases include but are not limited to:Hashimoto's thyroiditis, Grave's disease, multiple sclerosis, perniciousanemia, Addison's disease, insulin-dependent diabetes mellitus,rheumatoid arthritis, systemic lupus erythematosus (SLE or lupus),dermatomyositis, Crohn's disease, Wegener's granulomatosis, AntiGlomerular Basement Membrane Disease, Antiphospholipid Syndrome, 25Dermatitis Herpetiformis, Allergic Encephalomyelitis,Glomerulonephritis, Membranous Glomerulonephritis, Goodpasture Syndrome,Lambert-Eaton, Myasthenic Syndrome, Myasthenia Gravis, BullousPemphigoid, Polyendocrinopathies, Reiter's Disease, and Stiff-ManSyndrome.

Thus, this invention also provides a method for the treatment ofautoimmune diseases and conditions and the treatment of diseasesassociated with conformational unstable or misfolded proteins byadministering an effective amount of a compound of the presentinvention, to a subject, e.g. a mammal (and more particularly a human)in need of such treatment.

The compounds of the present invention can also be useful for thetreatment of diseases associated with conformational unstable ormisfolded proteins. Examples of diseases associated with conformationalunstable or misfolded proteins include but are not limited to: cysticfibrosis (CFTR), Marfan syndrom (fibrillin), Amyotrophic lateralsclerosis (superoxide dismutase), scurvy (collagen), maple syrup urinedisease (alpha-ketoacid dehydrogenase complex), osteogenesis imperfecta(typel procollagen pro-alpha), Creutzfeldt-Jakob disease (prion),Alzheimer's disease (beta-amyloid), familial amyloidosis (lysozyme),cataracts (crystallins), familial hypercholesterolemia (LDL receptor), αI-antitrypsin deficiency, Tay-Sachs disease (beta-hexosaminidase),retinitis pigmentosa (rhodopsin), and leprechaunism (insulin receptor).

Thus, this invention also provides a method for the treatment ofdiseases associated with conformational unstable or misfolded proteinsby administering an effective amount of a compound of the presentinvention, to a subject, e.g. a mammal (and more particularly a human)in need of such treatment.

In view of their useful pharmacological properties, the subjectcompounds may be formulated into various pharmaceutical forms foradministration purposes.

To prepare the pharmaceutical compositions of this invention, aneffective amount of a particular compound, in base or acid addition saltform, as the active ingredient is combined in intimate admixture with apharmaceutically acceptable carrier, which carrier may take a widevariety of forms depending on the form of preparation desired foradministration. These pharmaceutical compositions are desirably inunitary dosage form suitable, preferably, for administration orally,rectally, percutaneously, or by parenteral injection. For example, inpreparing the compositions in oral dosage form, any of the usualpharmaceutical media may be employed, such as, for example, water,glycols, oils, alcohols and the like in the case of oral liquidpreparations such as suspensions, syrups, elixirs and solutions; orsolid carriers such as starches, sugars, kaolin, lubricants, binders,disintegrating agents and the like in the case of powders, pills,capsules and tablets.

Because of their ease in administration, tablets and capsules representthe most advantageous oral dosage unit form, in which case solidpharmaceutical carriers are obviously employed. For parenteralcompositions, the carrier will usually comprise sterile water, at leastin large part, though other ingredients, to aid solubility for example,may be included. Injectable solutions, for example, may be prepared inwhich the carrier comprises saline solution, glucose solution or amixture of saline and glucose solution. Injectable suspensions may alsobe prepared in which case appropriate liquid carriers, suspending agentsand the like may be employed. In the compositions suitable forpercutaneous administration, the carrier optionally comprises apenetration enhancing agent and/or a suitable wetting agent, optionallycombined with suitable additives of any nature in minor proportions,which additives do not cause a significant deleterious effect to theskin. Said additives may facilitate the administration to the skinand/or may be helpful for preparing the desired compositions. Thesecompositions may be administered in various ways, e.g., as a transdermalpatch, as a spot-on, as an ointment. It is especially advantageous toformulate the aforementioned pharmaceutical compositions in dosage unitform for ease of administration and uniformity of dosage. Dosage unitform as used in the specification and claims herein refers to physicallydiscrete units suitable as unitary dosages, each unit containing apredetermined quantity of active ingredient calculated to produce thedesired therapeutic effect in association with the requiredpharmaceutical carrier. Examples of such dosage unit forms are tablets(including scored or coated tablets), capsules, pills, powder packets,wafers, injectable solutions or suspensions, teaspoonfuls,tablespoonfuls and the like, and segregated multiples thereof.

It is especially advantageous to formulate the aforementionedpharmaceutical compositions in dosage unit form for ease ofadministration and uniformity of dosage. Dosage unit form as used in thespecification and claims herein refers to physically discrete unitssuitable as unitary dosages, each unit containing a predeterminedquantity of active ingredient, calculated to produce the desiredtherapeutic effect, in association with the required pharmaceuticalcarrier. Examples of such dosage unit forms are tablets (includingscored or coated tablets), capsules, pills, powder packets, wafers,injectable solutions or suspensions, teaspoonfuls, tablespoonfuls andthe like, and segregated multiples thereof.

The compound of the invention is administered in an amount sufficient toinhibit the interaction between MDM2 and p53 or other cellular proteinsthat induce apoptosis, induce cellular death, or regulate the cellcycle.

The oncogenic potential of MDM2 is not only determined by its ability tosuppress p53, but also by its ability to regulate other tumoursuppressor proteins, e.g. the retinoblastoma protein pRb and the closelyassociated E2F1 transcription factor.

Thus, the compound of the invention is administered in an amountsufficient to modulate the interaction between MDM2 and the E2Ftranscription factors.

Those skilled in the art could easily determine the effective amountfrom the test results presented hereinafter. In general it iscontemplated that a therapeutically effective amount would be from 0.005mg/kg to 100 mg/kg body weight, and in particular from 0.005 mg/kg to 10mg/kg body weight. It may be appropriate to administer the required doseas single, two, three, four or more sub-doses at appropriate intervalsthroughout the day. Said sub-doses may be formulated as unit dosageforms, for example, containing 0.5 to 500 mg, and in particular 10 mg to500 mg of active ingredient per unit dosage form.

As another aspect of the present invention, a combination of a p53-MDM2inhibitor with another anticancer agent is envisaged, especially for useas a medicine, more specifically in the treatment of cancer or relateddiseases.

For the treatment of the above conditions, the compounds of theinvention may be advantageously employed in combination with one or moreother medicinal agents, more particularly, with other anti-canceragents. Examples of anti-cancer agents are:

-   -   platinum coordination compounds for example cisplatin,        carboplatin or oxalyplatin;    -   taxane compounds for example paclitaxel or docetaxel;    -   topoisomerase I inhibitors such as camptothecin compounds for        example irinotecan or topotecan;    -   topoisomerase II inhibitors such as anti-tumour podophyllotoxin        derivatives for example etoposide or teniposide;    -   anti-tumour vinca alkaloids for example vinblastine, vincristine        or vinorelbine;    -   anti-tumour nucleoside derivatives for example 5-fluorouracil,        gemcitabine or capecitabine;    -   alkylating agents such as nitrogen mustard or nitrosourea for        example cyclophosphamide, chlorambucil, carmustine or lomustine;    -   anti-tumour anthracycline derivatives for example daunorubicin,        doxorubicin, idarubicin or mitoxantrone;    -   HER2 antibodies for example trastuzumab;    -   estrogen receptor antagonists or selective estrogen receptor        modulators for example tamoxifen, toremifene, droloxifene,        faslodex or raloxifene;    -   aromatase inhibitors such as exemestane, anastrozole, letrazole        and vorozole;    -   differentiating agents such as retinoids, vitamin D and retinoic        acid metabolism blocking agents (RAMBA) for example accutane;    -   DNA methyl transferase inhibitors for example azacytidine;    -   kinase inhibitors for example flavoperidol, imatinib mesylate or        gefitinib;    -   farnesyltransferase inhibitors;    -   HDAC inhibitors;    -   other inhibitors of the ubiquitin-proteasome pathway for example        Velcade; or    -   Yondelis.

The term “platinum coordination compound” is used herein to denote anytumour cell growth inhibiting platinum coordination compound whichprovides platinum in the form of an ion.

The term “taxane compounds” indicates a class of compounds having thetaxane ring system and related to or derived from extracts from certainspecies of yew (Taxus) trees.

The term “topisomerase inhibitors” is used to indicate enzymes that arecapable of altering DNA topology in eukaryotic cells. They are criticalfor important cellular functions and cell proliferation. There are twoclasses of topoisomerases in eukaryotic cells, namely type I and typeII. Topoisomerase I is a monomeric enzyme of approximately 100,000molecular weight. The enzyme binds to DNA and introduces a transientsingle-strand break, unwinds the double helix (or allows it to unwind)and subsequently reseals the break before dissociating from the DNAstrand. Topisomerase II has a similar mechanism of action which involvesthe induction of DNA strand breaks or the formation of free radicals.

The term “camptothecin compounds” is used to indicate compounds that arerelated to or derived from the parent camptothecin compound which is awater-insoluble alkaloid derived from the Chinese tree Camptothecinacuminata and the Indian tree Nothapodytes foetida.

The term “podophyllotoxin compounds” is used to indicate compounds thatare related to or derived from the parent podophyllotoxin, which isextracted from the mandrake plant.

The term “anti-tumour vinca alkaloids” is used to indicate compoundsthat are related to or derived from extracts of the periwinkle plant(Vinca rosea).

The term “alkylating agents” encompass a diverse group of chemicals thathave the common feature that they have the capacity to contribute, underphysiological conditions, alkyl groups to biologically vitalmacromolecules such as DNA. With most of the more important agents suchas the nitrogen mustards and the nitrosoureas, the active alkylatingmoieties are generated in vivo after complex degradative reactions, someof which are enzymatic. The most important pharmacological actions ofthe alkylating agents are those that disturb the fundamental mechanismsconcerned with cell proliferation in particular DNA synthesis and celldivision. The capacity of alkylating agents to interfere with DNAfunction and integrity in rapidly proliferating tissues provides thebasis for their therapeutic applications and for many of their toxicproperties.

The term “anti-tumour anthracycline derivatives” comprise antibioticsobtained from the fungus Strep. peuticus var. caesius and theirderivatives, characterised by having a tetracycline ring structure withan unusual sugar, daunosamine, attached by a glycosidic linkage.

Amplification of the human epidermal growth factor receptor 2 protein(HER 2) in primary breast carcinomas has been shown to correlate with apoor clinical prognosis for certain patients. Trastuzumab is a highlypurified recombinant DNA-derived humanized monoclonal IgG1 kappaantibody that binds with high affinity and specificity to theextracellular domain of the HER2 receptor.

Many breast cancers have estrogen receptors and growth of these tumourscan be stimulated by estrogen. The terms “estrogen receptor antagonists”and “selective estrogen receptor modulators” are used to indicatecompetitive inhibitors of estradiol binding to the estrogen receptor(ER). Selective estrogen receptor modulators, when bound to the ER,induces a change in the three-dimensional shape of the receptor,modulating its binding to the estrogen responsive element (ERE) on DNA.

In postmenopausal women, the principal source of circulating estrogen isfrom conversion of adrenal and ovarian androgens (androstenedione andtestosterone) to estrogens (estrone and estradiol) by the aromataseenzyme in peripheral tissues. Estrogen deprivation through aromataseinhibition or inactivation is an effective and selective treatment forsome postmenopausal patients with hormone-dependent breast cancer.

The term “antiestrogen agent” is used herein to include not onlyestrogen receptor antagonists and selective estrogen receptor modulatorsbut also aromatase inhibitors as discussed above.

The term “differentiating agents” encompass compounds that can, invarious ways, inhibit cell proliferation and induce differentiation.Vitamin D and retinoids are known to play a major role in regulatinggrowth and differentiation of a wide variety of normal and malignantcell types. Retinoic acid metabolism blocking agents (RAMBA's) increasethe levels of endogenous retinoic acids by inhibiting the cytochromeP450-mediated catabolism of retinoic acids.

DNA methylation changes are among the most common abnormalities in humanneoplasia. Hypermethylation within the promotors of selected genes isusually associated with inactivation of the involved genes. The term“DNA methyl transferase inhibitors” is used to indicate compounds thatact through pharmacological inhibition of DNA methyl transferase andreactivation of tumour suppressor gene expression.

The term “kinase inhibitors” comprises potent inhibitors of kinases thatare involved in cell cycle progression and programmed cell death(apoptosis).

The term “farnesyltransferase inhibitors” is used to indicate compoundsthat were designed to prevent farnesylation of Ras and otherintracellular proteins. They have been shown to have effect on malignantcell proliferation and survival.

The term “histone deacetylase inhibitor” or “inhibitor of histonedeacetylase” is used to identify a compound, which is capable ofinteracting with a histone deacetylase and inhibiting its activity, moreparticularly its enzymatic activity. Inhibiting histone deacetylaseenzymatic activity means reducing the ability of a histone deacetylaseto remove an acetyl group from a histone.

The term “other inhibitors of the ubiquitin-proteasome pathway” is usedto identify compounds that inhibit the targeted destruction of cellularproteins in the proteasome, including cell cycle regulatory proteins.

As stated above, the compounds of the present invention also havetherapeutic applications in sensitising tumour cells for chemotherapyand radiotherapy.

Hence the compounds of the present invention can be used as“radiosensitizer” and/or “chemosensitizer” or can be given incombination with another “radiosensitizer” and/or “chemosensitizer”.

The term “radiosensitizer”, as used herein, is defined as a molecule,preferably a low molecular weight molecule, administered to animals intherapeutically effective amounts to increase the sensitivity of thecells to ionizing radiation and/or to promote the treatment of diseaseswhich are treatable with ionizing radiation.

The term “chemosensitizer”, as used herein, is defined as a molecule,preferably a low molecular weight molecule, administered to animals intherapeutically effective amounts to increase the sensitivity of cellsto chemotherapy and/or promote the treatment of diseases which aretreatable with chemotherapeutics.

Several mechanisms for the mode of action of radiosensitizers have beensuggested in the literature including: hypoxic cell radiosensitizers(e.g., 2-nitroimidazole compounds, and benzotriazine dioxide compounds)mimicking oxygen or alternatively behave like bioreductive agents underhypoxia; non-hypoxic cell radiosensitizers (e.g., halogenatedpyrimidines) can be analogues of DNA bases and preferentiallyincorporate into the DNA of cancer cells and thereby promote theradiation-induced breaking of DNA molecules and/or prevent the normalDNA repair mechanisms; and various other potential mechanisms of actionhave been hypothesized for radiosensitizers in the treatment of disease.

Many cancer treatment protocols currently employ radiosensitizers inconjunction with radiation of x-rays. Examples of x-ray activatedradiosensitizers include, but are not limited to, the following:metronidazole, misonidazole, desmethylmisonidazole, pimonidazole,etanidazole, nimorazole, mitomycin C, RSU 1069, SR 4233, EO9, RB 6145,nicotinamide, 5-bromodeoxyuridine (BUdR), 5-iododeoxyuridine (IUdR),bromodeoxycytidine, fluorodeoxyuridine (FudR), hydroxyurea, cisplatin,and therapeutically effective analogs and derivatives of the same.

Photodynamic therapy (PDT) of cancers employs visible light as theradiation activator of the sensitizing agent. Examples of photodynamicradiosensitizers include the following, but are not limited to:hematoporphyrin derivatives, Photofrin, benzoporphyrin derivatives, tinetioporphyrin, pheoborbide-a, bacteriochlorophyll-a, naphthalocyanines,phthalocyanines, zinc phthalocyanine, and therapeutically effectiveanalogs and derivatives of the same.

Radiosensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds which promote the incorporationof radiosensitizers to the target cells; compounds which control theflow of therapeutics, nutrients, and/or oxygen to the target cells;chemotherapeutic agents which act on the tumour with or withoutadditional radiation; or other therapeutically effective compounds fortreating cancer or other disease.

Chemosensitizers may be administered in conjunction with atherapeutically effective amount of one or more other compounds,including but not limited to: compounds which promote the incorporationof chemosensitizers to the target cells; compounds which control theflow of therapeutics, nutrients, and/or oxygen to the target cells;chemotherapeutic agents which act on the tumour or other therapeuticallyeffective compounds for treating cancer or other disease.

In view of their useful pharmacological properties, the components ofthe combinations according to the invention, i.e. the other medicinalagent and the p53-MDM inhibitor may be formulated into variouspharmaceutical forms for administration purposes. The components may beformulated separately in individual pharmaceutical compositions or in aunitary pharmaceutical composition containing both components.

The present invention therefore also relates to a pharmaceuticalcomposition comprising the other medicinal agent and the p53-MDMinhibitor together with one or more pharmaceutical carriers.

The present invention further relates to the use of a combinationaccording to the invention in the manufacture of a pharmaceuticalcomposition for inhibiting the growth of tumour cells.

The present invention further relates to a product containing as firstactive ingredient a p53-MDM2 inhibitor according to the invention and assecond active ingredient an anticancer agent, as a combined preparationfor simultaneous, separate or sequential use in the treatment ofpatients suffering from cancer.

The other medicinal agent and p53-MDM2 inhibitor may be administeredsimultaneously (e.g. in separate or unitary compositions) orsequentially in either order. In the latter case, the two compounds willbe administered within a period and in an amount and manner that issufficient to ensure that an advantageous or synergistic effect isachieved. It will be appreciated that the preferred method and order ofadministration and the respective dosage amounts and regimes for eachcomponent of the combination will depend on the particular othermedicinal agent and p53-MDM2 inhibitor being administered, their routeof administration, the particular tumour being treated and theparticular host being treated. The optimum method and order ofadministration and the dosage amounts and regime can be readilydetermined by those skilled in the art using conventional methods and inview of the information set out herein.

The platinum coordination compound is advantageously administered in adosage of 1 to 500 mg per square meter (mg/m²) of body surface area, forexample 50 to 400 mg/m², particularly for cisplatin in a dosage of about75 mg/m² and for carboplatin in about 300 mg/m² per course of treatment.

The taxane compound is advantageously administered in a dosage of 50 to400 mg per square meter (mg/m²) of body surface area, for example 75 to250 mg/m², particularly for paclitaxel in a dosage of about 175 to 250mg/m² and for docetaxel in about 75 to 150 mg/m² per course oftreatment.

The camptothecin compound is advantageously administered in a dosage of0.1 to 400 mg per square meter (mg/m²) of body surface area, for example1 to 300 mg/m², particularly for irinotecan in a dosage of about 100 to350 mg/m² and for topotecan in about 1 to 2 mg/m² per course oftreatment.

The anti-tumour podophyllotoxin derivative is advantageouslyadministered in a dosage of 30 to 300 mg per square meter (mg/m²) ofbody surface area, for example 50 to 250 mg/m², particularly foretoposide in a dosage of about 35 to 100 mg/m² and for teniposide inabout 50 to 250 mg/m² per course of treatment.

The anti-tumour vinca alkaloid is advantageously administered in adosage of 2 to 30 mg per square meter (mg/m²) of body surface area,particularly for vinblastine in a dosage of about 3 to 12 mg/m², forvincristine in a dosage of about 1 to 2 mg/m², and for vinorelbine indosage of about 10 to 30 mg/m² per course of treatment.

The anti-tumour nucleoside derivative is advantageously administered ina dosage of 200 to 2500 mg per square meter (mg/m²) of body surfacearea, for example 700 to 1500 mg/m², particularly for 5-FU in a dosageof 200 to 500 mg/m², for gemcitabine in a dosage of about 800 to 1200mg/m² and for capecitabine in about 1000 to 2500 mg/m² per course oftreatment.

The alkylating agents such as nitrogen mustard or nitrosourea isadvantageously administered in a dosage of 100 to 500 mg per squaremeter (mg/m²) of body surface area, for example 120 to 200 mg/m²,particularly for cyclophosphamide in a dosage of about 100 to 500 mg/m²,for chlorambucil in a dosage of about 0.1 to 0.2 mg/kg, for carmustinein a dosage of about 150 to 200 mg/m², and for lomustine in a dosage ofabout 100 to 150 mg/m² per course of treatment.

The anti-tumour anthracycline derivative is advantageously administeredin a dosage of 10 to 75 mg per square meter (mg/m²) of body surfacearea, for example 15 to 60 mg/m², particularly for doxorubicin in adosage of about 40 to 75 mg/m², for daunorubicin in a dosage of about 25to 45 mg/m², and for idarubicin in a dosage of about 10 to 15 mg/m² percourse of treatment.

Trastuzumab is advantageously administered in a dosage of 1 to 5 mg persquare meter (mg/m²) of body surface area, particularly 2 to 4 mg/m² percourse of treatment.

The antiestrogen agent is advantageously administered in a dosage ofabout 1 to 100 mg daily depending on the particular agent and thecondition being treated. Tamoxifen is advantageously administered orallyin a dosage of 5 to 50 mg, preferably 10 to 20 mg twice a day,continuing the therapy for sufficient time to achieve and maintain atherapeutic effect. Toremifene is advantageously administered orally ina dosage of about 60 mg once a day, continuing the therapy forsufficient time to achieve and maintain a therapeutic effect.Anastrozole is advantageously administered orally in a dosage of about 1mg once a day. Droloxifene is advantageously administered orally in adosage of about 20-100 mg once a day. Raloxifene is advantageouslyadministered orally in a dosage of about 60 mg once a day. Exemestane isadvantageously administered orally in a dosage of about 25 mg once aday.

These dosages may be administered for example once, twice or more percourse of treatment, which may be repeated for example every 7, 14, 21or 28 days.

The compounds of formula (I), the pharmaceutically acceptable acidaddition salts and stereoisomeric forms thereof can have valuablediagnostic properties in that they can be used for detecting oridentifying an p53-MDM2 interaction in a biological sample comprisingdetecting or measuring the formation of a complex between a labelledcompound and/or p53 and/or MDM2 and or other molecules, peptides,proteins, enzymes or receptors.

The detecting or identifying methods can use compounds that are labelledwith labelling agents such as radioisotopes, enzymes, fluorescentsubstances, luminous substances, etc. Examples of the radioisotopesinclude ¹²⁵I, ¹³¹I, ³H and ¹⁴C. Enzymes are usually made detectable byconjugation of an appropriate substrate which, in turn catalyses adetectable reaction. Examples thereof include, for example,beta-galactosidase, beta-glucosidase, alkaline phosphatase, peroxidaseand malate dehydrogenase, preferably horseradish peroxidase. Theluminous substances include, for example, luminol, luminol derivatives,luciferin, acquorin and luciferase. Biological samples can be defined asbody tissue or body fluids. Examples of body fluids are cerebrospinalfluid, blood, plasma, serum, urine, sputum, saliva and the like.

The following examples illustrate the present invention.

EXPERIMENTAL PART

Hereinafter, “DMF” is defined as N,N-dimethylformamide, “DCM” is definedas dichloromethane, “DIPE” is defined as diisopropyl ether, “EtOAc” isdefined as ethyl acetate, “EtOH” is defined as ethanol, “EDC” is definedas N′-(ethylcarbonimidoyl)-N,N-dimethyl-1,3-propanediamine,monohydrochloride, “MeOH” is defined as methanol, “THF” is defined astetrahydrofuran., “HOBT” is defined as 1-hydroxybenzotriazole.

A. Preparation of the Intermediate Compounds Example A1 a) Preparationof Intermediate 1

A mixture of 1-fluoro-4-nitro-benzene (0.0142 mol),1H-indole-3-ethanamine (0.0129 mol) andN-ethyl-N-(1-methylethyl)-2-propanamine (0.032 mol) was stirred at 210°C. for 18 hours, then brought to room temperature, and decanted. Theresidue was taken up in acetonitrile/water. The precipitate wasfiltered, washed with diethyl ether and dried, yielding 2.3 g (64%) ofintermediate 1.

b) Preparation of Intermediate 2

A mixture of intermediate 1 (0.0078 mol) and Raney Nickel (2.2 g) inEtOH (50 ml) was hydrogenated at room temperature for 3 hours under a 3bar pressure, then filtered over celite. Celite was washed withDCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM,dried (MgSO₄), filtered, and the solvent was evaporated, yielding 1.88 g(95%) of intermediate 2.

Example A2 a) Preparation of Intermediate 3

A mixture of 2-ethoxy-1-methoxy-4-nitro-benzene (0.009 mol),1H-indole-3-ethanamine (0.009 mol) andN-ethyl-N-(1-methylethyl)-2-propanamine (0.0228 mol) was stirred at 210°C. for 24 hours, then brought to room temperature, taken up in DCM/MeOHand dried. The residue was taken up in DCM (few) and purified by columnchromatography over silica gel (35-70 μm) (eluent: cyclohexane/DCM30/70). The pure fractions were collected and the solvent wasevaporated, yielding 0.6 g (20%) of intermediate 3.

b) Preparation of Intermediate 4

A mixture of intermediate 3 (0.002 mol) and H₂/Raney Nickel (0.6 g) inMeOH (100 ml) was hydrogenated at room temperature for 1 hour and 30minutes under a 3 bar pressure, then filtered over celite. Celite waswashed with DCM/MeOH. The filtrate was evaporated. The residue was takenup in DCM/MeOH (few), dried (MgSO₄), filtered and the solvent wasevaporated, yielding 0.45 g (83%) of intermediate 4.

Example A3 a) Preparation of Intermediate 5

A mixture of N-3-pyridinyl-acetamide (0.038 mol),1-fluoro-4-nitro-benzene (0.05 mol), copper(I)chloride (0.0038 mol) andpotassium carbonate (0.076 mol) in xylene (60 ml) was stirred andrefluxed for 18 hours, then brought to room temperature. Water wasadded. The mixture was filtered over celite. Celite was washed with DCM.The filtrate was evaporated. The residue was purified by columnchromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH₄OH97/3/0.1). The pure fractions were collected and the solvent wasevaporated, yielding 6.4 g (65%) of intermediate 5.

b) Preparation of Intermediate 6

Sodium hydroxide (concentrated) (10 ml) was added to a mixture ofintermediate 5 (0.025 mol) in EtOH (80 ml). The mixture was stirred andrefluxed for 2 hours, then brought to room temperature. Water was added.The mixture was stirred for 15 minutes then filtered. The residue waspurified by column chromatography over silica gel (70-200 μm) (eluent:DCM/MeOH 100/0 to 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 1.5 g (28%) of intermediate 6.

c) Preparation of Intermediate 7

A mixture of intermediate 6 (0.007 mol) and Raney Nickel (1.5 g) in MeOH(30 ml) and THF (10 ml) was hydrogenated at room temperature for 1 hourunder a 3 bar pressure, then filtered over celite. Celite was washedwith DCM/MeOH. The filtrate was evaporated. The residue was taken up inDCM. The organic layer was separated, dried (MgSO₄), filtered, and thesolvent was evaporated, yielding 1.2 g (92%) of intermediate 7.

Example A4 Preparation of Intermediate 8

1H-indole-3-propanoic acid (0.0264 mol) then 1-hydroxybenzotriazole(0.0344 mol) thenN′-(ethylcarbonimidoyl)-N,N-dimethyl-1,3-propanediamine,monohydrochloride (=EDCI) (0.0344 mol) were added to a mixture of1,4-benzenediamine (0.137 mol) in THF (200 ml) and DCM (200 ml) under N₂flow. The mixture was stirred at room temperature for 24 hours, pouredout into water and extracted with DCM. The organic layer was separated,dried (MgSO₄), filtered, and the solvent was evaporated. The residue waspurified by column chromatography over silica gel (20-45 μm) (eluent:DCM/MeOH/NH₄OH 97/3/0.5). The pure fractions were collected and thesolvent was evaporated, yielding 1.75 g (24%) of intermediate 8.

Example A5 a) Preparation of Intermediate 9

A mixture of 1-fluoro-4-nitro-benzene (0.0025 mol),1-methyl-1H-indole-3-ethanamine (0.0023 mol) andN-ethyl-N-(1-methylethyl)-2-propanamine (0.0057 mol) was stirred at 200°C. for 2 hours, then brought to room temperature. Water and DCM wereadded. The organic layer was separated, dried (MgSO₄), filtered, and thesolvent was evaporated. The residue (0.8 g) was purified by columnchromatography over silica gel (35-70 μm) (eluent: DCM 100). The purefractions were collected and the solvent was evaporated. The residue(0.45 g) was taken up in DIPE. The precipitate was filtered off anddried, yielding 0.33 g (66%) of intermediate 9.

b) Preparation of Intermediate 10

A mixture of intermediate 9 (0.0011 mol) and Raney Nickel (0.4 g) inMeOH (20 ml) was hydrogenated at room temperature for 1 hour under a 3bar pressure, then filtered over celite. Celite was washed withDCM/MeOH. The filtrate was evaporated. The residue was taken up in DCM.The organic layer was separated, dried (MgSO₄), filtered, and thesolvent was evaporated, yielding 0.305 g (97%) of intermediate 10.

Example A6 a) Preparation of Intermediate 11

A mixture of 4-fluoro-benzonitrile (0.071 mol), 1H-indole-3-ethanamine(0.071 mol) and N-ethyl-N-(1-methylethyl)-2-propanamine (0.1775 mol) wasstirred at 210° C. for 16 hours, then brought to room temperature andtaken up in DCM/MeOH. The organic layer was washed with HCl 3N, dried(MgSO₄), filtered and the solvent was evaporated. The residue was takenup in diethyl ether/acetonitrile. The precipitate was filtered off anddried, yielding 8.07 g (43%) of intermediate 11, melting point 144° C.

b) Preparation of Intermediate 12

A mixture of intermediate 11 (0.0115 mol) and sodium hydroxide (0.17mol) in EtOH (50 ml) and water (50 ml) was stirred and refluxed for 18hours, then brought to room temperature. The solvent was evaporated. Theresidue was taken up in sodium hydroxide 3N. The aqueous layer waswashed with DCM and acidified till pH 5 was obtained. The precipitatewas filtered off and dried, yielding 1.06 g (35%) of intermediate 12,melting point 225° C.

c) Preparation of Intermediate 13

A mixture of intermediate 12 (0.0037 mol), 4-pyridinamine (0.0037 mol),2-chloro-1-methyl-pyridinium, iodide (0.0113 mol) and triethylamine(0.015 mol) in acetonitrile (100 ml) was stirred and refluxed for 90minutes, then brought to room temperature. The solvent was evaporated.The residue was taken up in DCM/MeOH. The organic layer was washed withpotassium carbonate 10%, dried (MgSO₄), filtered and the solvent wasevaporated till dryness. The residue was purified by flash columnchromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH₄OH95/5/0.1). Two fractions were collected and the solvent was evaporated,yielding 0.06 g F1 and 0.08 g F2. F1 was crystallized from diethylether/acetonitrile. The precipitate was filtered off and dried, yieldinga first batch of 0.032 g (2.4%) of intermediate 13. F2 and the motherlayer were combined and crystallized from diethyl ether/acetonitrile.The precipitate was filtered off and dried, yielding a second batch of0.105 g (10%) of intermediate 13, melting point 200° C.

Example A7 Preparation of Intermediate 14

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (227 mg, 0.0015 mol) and withaddition of triethylamine (0.22 ml). After workup, the residue waspurified by column chromatography over silica gel (40-63 μm) (eluent:EtOAc). The pure fractions were collected and the solvent wasevaporated, yielding 256 mg (52%) of intermediate 14 as a pale yellowoil.

Example A8 Preparation of Intermediate 15

Similar procedure as method 4 (see Example A22/22) was followed,starting from benzyl 1-piperazinecarboxylate (1.3 ml, 0.0069 mol) and4-chloro-2-pyridinemethanol (500 mg, 0.0034 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 840 mg (70%) of intermediate 15 as anorange oil.

Example A9 Preparation of Intermediate 16

Similar procedure as method 4 (see Example A22/22) was followed,starting from 4-amino-butan-1-ol (310 mg, 0.0021 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 85/15). The pure fractions were collected and thesolvent was evaporated, yielding 55 mg (17%) of intermediate 16 as ayellow oil.

Example A10 Preparation of Intermediate 17

Similar procedure as method 5 (see Example A22/34) was followed,starting from 4-(2-amino-ethyl)-piperazine-1-carboxylic acid tert-butylester (579 mg, 0.0025 mol) and 4-chloro-2-pyridinecarboxaldehyde (325mg, 0.0023 mol). After workup, the residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 90/10). Thepure fractions were collected and the solvent was evaporated, yielding425 mg (53%) of intermediate 17 as a yellow oil.

Example A11 Preparation of Intermediate 18

Similar procedure as method 5 (see Example A22/34) was followed,starting from 3-amino-propionic acid methyl ester hydrochloride (351 mg,0.0025 mol) and 4-chloro-2-pyridinecarboxaldehyde (325 mg, 0.0023 mol)and with addition of triethylamine (0.35 ml, 0.0025 mol). After workup,the residue was purified by column chromatography over silica gel (40-63μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 160 mg (30%) of intermediate 18 as ayellow oil.

Example A12 Preparation of Intermediate 19

Similar procedure as method 3 (see Example A22/20) was followed,starting from bromo-acetic acid methyl ester (0.26 ml, 0.0021 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: AcOEt/cyclohexane 60/40). The pure fractions were collected andthe solvent was evaporated, yielding 170 mg (38%) of intermediate 19 asa yellow oil.

Example A13 Preparation of Intermediate 20

Method 6

4-Amino-1-butanol (0.13 ml, 0.0014 mol) was added at room temperature toa mixture of 1-(4-chloro-2-pyridinyl)-ethanone (200 mg, 0.0013 mol),para-toluene sulfonic acid (123 mg, 0.00065 mol), and 3 Å molecularsieves in MeOH (4 ml). The mixture was stirred 6 hours at roomtemperature, cooled down to 0° C., and sodium borohydride (98 mg, 0.0026mol) was slowly added. The mixture was stirred at room temperature for18 hours. Molecular sieves were filtered off, and the mixture was pouredout into water and the solvent was evaporated. The aqueous layer wasbasified with a saturated solution of sodium hydrogen carbonate, andextracted 3 times with DCM. The organic layer was separated, washed withbrine, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 269 mg (91%) of intermediate 20 as ayellow oil.

Example A14 Preparation of Intermediate 21

A mixture of α-(phenylmethyl)-1H-indole-3-acetic acid (94 mg, 0.00035mol) and 1,1 carbonyldiimidazole (59 mg, 0.00036 mol, added portionwise)in DCM (1 ml) was stirred 3 hours at room temperature under argon.N,O-dimethylhydroxylamine hydrochloride (36 mg, 0.00037 mol) was added,and the mixture was stirred 3 more hours at room temperature, cooleddown to 0° C., then poured out into water. pH was adjusted to 10 with a4N solution of sodium hydroxide, and aqueous layer was extracted withEtOAc. The organic layer was separated, washed with a 3N solution ofhydrochloric acid, dried (MgSO₄), filtered, and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The purefractions were collected and the solvent was evaporated, yielding 52 mg(47%) of intermediate 21.

Example A15 a) Preparation of Intermediate 22

To a solution of intermediate 1 (3.0 g, 0.011 mol) in DCM (130 ml), wasadded 4-dimethylaminopyridine (261 mg, 0.0021 mol) anddi-tert-butyldicarbonate (14.0 g, 0.064 mol). The mixture was stirred atroom temperature for 5 hours. The reaction was quenched by addition ofwater and extracted twice with DCM. The organic layer was washedsuccessively with a saturated solution of sodium bicarbonate and withbrine, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc/cyclohexane 10/90 to 20/80). The pure fractions werecollected and the solvent was evaporated, yielding 4.65 g (90%) ofintermediate 22 as a yellow solid.

b) Preparation of Intermediate 23

Raney nickel (3 g) was added to a solution of intermediate 22 (4.7 g,0.0097 mol) in ethanol (15 ml) and THF (15 ml). The reaction mixture wasstirred under 1 atmosphere of hydrogen for 16 hours. To complete thereaction, raney nickel (1 g) was added and the mixture was stirred under1 atmosphere of hydrogen for 4 more hours. The mixture was filteredthrough a celite pad and the solvent was evaporated. The residue waspurified by column chromatography over silica gel (40-63 μm) (eluent:EtOAc/cyclohexane 10/90 to 20/80). The pure fractions were collected andthe solvent was evaporated, yielding 4.0 g (92%) of intermediate 23 as ayellow foam.

Example A16 a) Preparation of Intermediate 24

Bromine (0.0104 mol) then a solution of sodium nitrite (0.0362 mol) inwater (3 ml) were added drop wise at −10° C. to a mixture of6,7-dihydro-5H-1-pyridin-4-amine (0.0112 mol) in aqueous hydrogenbromide (48%) (5 ml). The mixture was brought back to 20° C. Ice wasadded. The mixture was basified with concentrated sodium hydroxide andextracted with EtOAc. The organic layer was separated, dried (MgSO₄),filtered, and the solvent was evaporated, yielding: 2 g (90%) ofintermediate 24.

b) Preparation of Intermediate 25

Meta-chloroperbenzoic acid (0.012 mol) was added to a mixture ofintermediate 24 (0.01 mol) in DCM (15 ml). The mixture was stirred atroom temperature for 12 hours. Sodium hydroxide 3N and water were added.The mixture was extracted three times with DCM. The organic layer waswashed with water, dried (MgSO₄), filtered, and the solvent wasevaporated, yielding 1.85 g (86%) of intermediate 25.

c) Preparation of Intermediate 26

A mixture of intermediate 25 (0.0086 mol) in acetic anhydride (18 ml)was stirred at 100° C. for 30 minutes, then cooled to room temperatureand evaporated. The residue was taken up in NaHCO₃ and EtOAc andfiltered over celite. Celite was washed with EtOAc. The organic layerwas separated, dried (MgSO₄), filtered, and the solvent was evaporated,yielding 1.63 g (73%) of intermediate 26.

d) Preparation of Intermediate 27

A mixture of intermediate 26 (0.0074 mol) in MeOH (10 ml) and sodiumhydroxide 3N (80 ml) was stirred at room temperature for 30 minutes,then stirred at 80° C. for 10 minutes then brought back to roomtemperature. MeOH was evaporated. The mixture was extracted twice withDCM then washed with saturated NaCl. The organic layer was separated,dried (MgSO₄), filtered, and the solvent was evaporated, yielding 1.02 g(64%) of intermediate 27.

Example A17 a) Preparation of Intermediate 28

Bromine (1.3 ml) then a solution of sodium nitrite (3.3 g) in water (4ml) were added drop wise at −10° C. to a solution of5,6,7,8-tetrahydro-4-quinolinamine (0.0135 mol) in aqueous hydrogenbromide (48%) (6.7 ml). The mixture was brought back to 20° C., pouredout on ice, basified with concentrated sodium hydroxide and extractedwith EtOAc. The organic layer was separated, dried (MgSO₄), filtered,and the solvent was evaporated, yielding 2.2 g (77%) of intermediate 28.

b) Preparation of Intermediate 29

Meta-chloroperbenzoic acid (0.0125 mol) was added to a mixture ofintermediate 28 (0.0104 mol) in DCM (20 ml). The mixture was stirred atroom temperature for 12 hours. Sodium hydroxide 3N and ice were added.The mixture was extracted twice with DCM. The organic layer was dried(MgSO₄), filtered, and the solvent was evaporated, yielding 3 g (100%)of intermediate 29.

c) Preparation of Intermediate 30

A mixture of intermediate 29 (0.0086 mol) in acetic anhydride (22 ml)was stirred at 100° C. for 30 minutes, then cooled to room temperatureand evaporated. The residue was taken up in saturated NaHCO₃ and EtOAc.The mixture was stirred for 30 minutes. The organic layer was separated,dried (MgSO₄), filtered, and the solvent was evaporated, yielding 3.4 g(100%) of intermediate 30.

d) Preparation of Intermediate 31

A mixture of intermediate 30 (0.0104 mol) in MeOH (18 ml) and sodiumhydroxide 3N (150 ml) was stirred at room temperature for 30 minutes,then stirred at 80° C. for 10 minutes. MeOH was evaporated. The mixturewas extracted twice with DCM. The organic layer was washed withsaturated NaCl, dried (MgSO₄), filtered and the solvent was evaporated,yielding 1.84 g (77%) of intermediate 31.

Example A18 a) Preparation of Intermediate 32

A mixture of 2-ethoxy-4-nitroanisole (0.0107 mol), 6-methoxytryptamine(0.0107 mol) and diisopropylethylamine (0.0268 mol) was stirred at 210°C. for 5 hours then poured out on ice and extracted with DCM. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue was purified by column chromatography oversilica gel (15-40 μm) (eluent: DCM 100%). The pure fractions werecollected and the solvent was evaporated, yielding 0.85 g (22%) ofintermediate 32.

b) Preparation of Intermediate 33

A mixture of intermediate 32 (0.0023 mol) and Raney Nickel (0.85 g) inMeOH (42 ml) and THF (42 ml) was hydrogenated at room temperature for 2hours under a 3 bar pressure, then filtered over celite. The filtratewas evaporated, yielding 0.74 g (95%) of intermediate 33.

Example A19 a) Preparation of Intermediate 34

Oxalyl chloride (0.012 mol) was added drop wise at 0° C. to a solutionof 5-cyanoindole (0.007 mol) in diethyl ether (21 ml). The mixture wasstirred at 0° C. for 5 hours, then stirred at room temperatureovernight. The precipitate was filtered, washed with diethyl ether anddried, yielding 1.454 g of (73%) of intermediate 34.

b) Preparation of Intermediate 35

A solution of intermediate 34 (0.0027 mol) in DCM (12 ml) was added dropwise at 5° C. to a solution of N-Pyridin-4-yl-benzene-1,4-diamine (0.022mol) and N,N-diisopropylethylamine (0.0034 mol) in DCM (4 ml). Themixture was stirred and refluxed for a weekend, then cooled to roomtemperature. The precipitate was filtered off and dried. The residue wascrystallized from iPrOH. The precipitate was filtered off and dried,yielding 0.756 g of crude product. This fraction was purified by columnchromatography over kromasil (5 μm) (eluent: DCM/MeOH/NH₄OH 97/3/0.3 to87/13/1.3). The pure fractions were collected and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH 90/10/0.1 to 87/13/0.1).The pure fractions were collected and the solvent was evaporated,yielding 0.098 g (20%) of intermediate 35, melting point>264° C.

Example A20 a) Preparation of Intermediate 36

A mixture of 1H-benzimidazole-1-ethanamine (0.011 mol),1-fluoro-4-nitrobenzene (0.011 mol) and diisopropylethylamine (0.034mol) was stirred at 210° C. for 30 minutes. Diisopropylethylamine wasevaporated. The precipitate was dissolved in DCM/MeOH. The organic layerwas washed with potassium carbonate 10%, dried (MgSO₄), filtered and thesolvent was evaporated. The residue (3.2 g) was purified by columnchromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH98/2/0.5). The pure fractions were collected and the solvent wasevaporated. The residue (2.1 g, 77%) was crystallized from acetonitrile.The precipitate was filtered off and dried, yielding 1.3 g (47%) ofintermediate 36, melting point 144° C.

b) Preparation of Intermediate 37

A mixture of intermediate 36 (0.006 mol) and Raney Nickel (2 g) in MeOH(20 ml) was hydrogenated at room temperature under a 3 bar pressure,then filtered over celite. Celite was washed with DCM/MeOH. The filtratewas evaporated, yielding 1.7 g (100%) of intermediate 37.

Example A21 a) Preparation of Intermediate 38

A mixture of DL-Tryptophan, methyl ester (0.0078 mol),1-fluoro-4-nitrobenzene (0.0078 mol) and diisopropylethylamine (0.0353mol) was stirred at 210° C. for 4 hours, and then taken up in DCM/MeOH.HCl 3N was added. The mixture was stirred for 15 minutes. The organiclayer was washed with saturated NaHCO₃, dried (MgSO₄), filtered and thesolvent was evaporated. The residue was purified by columnchromatography over silica gel (35-70 μm) (eluent: DCM 100% thenDCM/MeOH 99/1). The pure fractions were collected and the solvent wasevaporated, yielding 0.75 g (28%) of intermediate 38.

b) Preparation of Intermediate 39

A mixture of intermediate 38 (0.0022 mol) and Raney nickel (0.75 g) inMeOH (100 ml) was hydrogenated at room temperature for 1 hour under a 3bar pressure, then filtered over celite. The filtrate was evaporated,yielding 0.65 g (96%) of intermediate 39.

c) Preparation of Intermediate 40

A mixture of intermediate 39 (0.139 mol) and 4-bromopyridinehydrochloride (0.139 mol) in acetic acid (450 ml) was stirred at 120° C.for 3 hours, poured out on ice, basified with concentrated sodiumhydroxide and extracted with DCM/MeOH (few). The organic layer wasseparated, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue (62.7 g) was purified by column chromatography over kromasil(20-45 μm) (eluent: DCM/MeOH/NH₄OH 93/7/0.5). The pure fractions werecollected and the solvent was evaporated, yielding 22 g (41%) ofintermediate 40.

d) Preparation of Intermediate 41

Lithium hydroxide, monohydrate (0.112 mol) was added portion wise at 0°C. to a solution of intermediate 40 (0.056 mol) in MeOH (86 ml) andwater (34.4 ml) under N₂ flow. The mixture was stirred at roomtemperature overnight, then evaporated till dryness, yielding: 22 g(quantitative yield) of intermediate 41.

Example A22 1) Preparation of Intermediate 42

A 2.5N solution of butyl lithium in hexane (3.4 ml, 0.0081 mol) wasadded to a solution of diisopropylamine (0.85 ml, 0.0088 mol) in THF (6ml) at −78° C. under Argon. The mixture was stirred 30 minutes at −78°C. 4-Chloro-3-methylpyridine hydrochloride (630 mg, 0.0038 mol) wasadded portionwise and the mixture was stirred 1 hour at −78° C. Diethylcarbonate (1.0 ml, 0.0096 mol) was added dropwise and the mixture wasstirred 1 more hour at −78° C., then warmed up to room temperature andlet stirred for 2.5 hours. The reaction was quenched by slow addition ofwater, and extracted twice with EtOAc. The organic layer was separated,washed with brine, dried (MgSO₄), filtered, and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: EtOAc/MeOH 100/0 then 90/10). The purefractions were collected and the solvent was evaporated, yielding 74 mg(9%) of intermediate 42.

2) Preparation of Intermediate 43

Triethyl phosphonoacetate (0.075 ml, 0.00038 mol) was added dropwise toa mixture of sodium hydride (10.6 mg, 0.00044 mol) in THF (5 ml) at roomtemperature under argon. The mixture was stirred at room temperature for20 minutes, then a solution of 4-chloro-3-pyridinecarboxaldehyde (50 mg,0.00035 mol) in THF (3 ml) was added dropwise. The mixture was stirredat room temperature for 16 hours, then poured out into water andextracted twice with EtOAc. The organic layer was separated, washed withbrine, dried (MgSO₄), filtered, and the solvent was evaporated, yielding77 mg (88%) of intermediate 43.

3) Preparation of Intermediate 44

A 2.5N solution of butyl lithium in hexane (0.80 ml, 0.0020 mol) wasadded to a solution of diisopropylamine (0.28 ml, 0.0020 mol) in THF (2ml) at −78° C. under Argon. The mixture was stirred 10 minutes at −78°C., then a solution of 4-chloropyridine (219 mg, 0.0019 mol) in THF (1ml) was added dropwise. The mixture was stirred 1.25 hour at −78° C.,then propionaldehyde (0.14 ml, 0.0019 mol) was added dropwise. Themixture was stirred 30 minutes at −78° C., and finally 4 hours at roomtemperature, poured out into water, and extracted with EtOAc. Theorganic layer was separated, washed with brine, dried (MgSO₄), filtered,and the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane90/10). The pure fractions were collected and the solvent wasevaporated, yielding 147 mg (44%) of intermediate 44.

4) Preparation of Intermediate 45

Method 2

A 3-Methyl magnesium bromide solution in diethyl ether (1.1 ml, 0.0032mol) was added to a-4-chloro-2-pyridinecarboxylic acid, methyl estersolution (200 mg, 0.0012 mol) in THF (4 ml) at −30° C. under Argon. Themixture was heated at 75° C. for 2 h 30, cooled down to 0° C. andquenched with water. The resulting mixture was made alkaline with asaturated solution of sodium hydrogen carbonate and extracted twice withEtOAc. The organic layer was washed with brine, dried (MgSO₄), filteredand the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane10/90). The pure fractions were collected and the solvent wasevaporated, yielding 54 mg (23%) of intermediate 45 as a brown oil.

5) Preparation of Intermediate 46

Methane sulfonyl chloride (98 μl, 0.0013 mol) was added dropwise to asolution of 4-chloro-2-pyridinemethanamine (150 mg, 0.0011 mol) andtriethylamine (177 μA, 0.0013 mol) in DCM (4 ml) at 0° C. under argon.The mixture was stirred at room temperature for 30 minutes. The reactionwas quenched with a saturated solution of sodium bicarbonate andextracted twice with DCM. The organic phase was dried (MgSO₄), filtered,and the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc). The purefractions were collected and the solvent was evaporated, yielding 82 mg(35%) of intermediate 46 as an orange oil.

6) Preparation of Intermediate 47

Method 7

Cyclopropanecarbonyl chloride (115 μl, 0.0013 mol) was added dropwise toa solution of 4-chloro-2-pyridinemethanamine (150 mg, 0.0011 mol) andtriethylamine (177 μA, 0.0013 mol) in DCM (4 ml) at 0° C. under Argon.The mixture was stirred at room temperature for 15 minutes. The reactionwas quenched with a saturated solution of sodium bicarbonate andextracted twice with DCM. The organic phase was dried (MgSO₄), filtered,and the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc). The purefractions were collected and the solvent was evaporated, yielding 85 mg(38%) of intermediate 47 as a white solid.

7) Preparation of Intermediate 48

Similar procedure as method 7 (see Example A22/6) was followed, startingfrom 4-chloro-2-pyridinemethanamine (150 mg, 0.0011 mol) andhydrocinnamoyl chloride (187 μl, 0.0013 mol). After workup, the residuewas purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc). The pure fractions were collected and the solvent wasevaporated, yielding 191 mg (66%) of intermediate 48 as a yellow solid.

8) Preparation of Intermediate 49

Similar procedure as method 7 (see Example A22/6) was followed, startingfrom 4-chloro-2-pyridinemethanamine (200 mg, 0.0014 mol) and propionylchloride (146 μl, 0.0017 mol). After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: EtOAc). Thepure fractions were collected and the solvent was evaporated, yielding126 mg (45%) of intermediate 49 as a colorless oil.

9) Preparation of Intermediate 50

Similar procedure as method 7 (sec Example A22/6) was followed, startingfrom 4-chloro-2-pyridinemethanamine (150 mg, 0.0011 mol) andphenylacetyl chloride (168 μl, 0.0013 mol). After workup, the residuewas purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc). The pure fractions were collected and the solvent wasevaporated, yielding 124 mg (45%) of intermediate 50 as a white solid.

10) Preparation of Intermediates 51 and 52

Method 1

To a solution of 4-chloro-2-pyridinecarboxaldehyde (377 mg, 0.0027 mol)in THF (4 ml) at 0° C. under Argon was added dropwise a 1.4Mcyclopropylmagnesium bromide solution in toluene/THF (75/25). Thereaction mixture was stirred at 0° C. for 1 hour, the dry ice bath wasremoved and the mixture was stirred at room temperature for 1 hour. Thereaction mixture was quenched by addition of water and extracted twicewith EtOAc. The organic layer was washed with brine, dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent:cyclohexane/EtOAc 80/20). The pure fractions were collected and thesolvent was evaporated, yielding 193 mg (39%) of intermediate 51 and 29mg of intermediate 52.

11) Preparation of Intermediate 53

Similar procedure as method 1 (see Example A22/10) was followed,starting from 4-chloro-2-pyridinecarboxaldehyde (300 mg, 0.0021 mol) anda 2.0M isopropylmagnesium chloride solution in THF (2.12 ml, 0.0042mol). After workup, the residue was purified by column chromatographyover silica gel (40-63 μm) (eluent: cyclohexane/EtOAc 80/20). The purefractions were collected and the solvent was evaporated, yielding 165 mg(42%) of intermediate 53 as a brown oil.

12) Preparation of Intermediate 54

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (298 mg, 0.0019 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: cyclohexane/EtOAc 90/10). The pure fractionswere collected and the solvent was evaporated, yielding 293 mg (62%) ofintermediate 54 as a yellow oil.

13) Preparation of Intermediate 55

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (100 mg, 0.00064 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: DCM/MeOH 85/15). The pure fractions werecollected and the solvent was evaporated, yielding 50 mg (45%) ofintermediate 55 as a yellow oil.

14) Preparation of Intermediate 56

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (100 mg, 0.00064 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: DCM/MeOH 90/10). The pure fractions werecollected and the solvent was evaporated, yielding 30 mg (25%) ofintermediate 56 as a yellow oil.

15) Preparation of Intermediate 57

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (157 mg, 0.0010 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions werecollected and the solvent was evaporated, yielding 134 mg (49%) ofintermediate 57 as a yellow oil.

16) Preparation of Intermediate 58

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (200 mg, 0.0013 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions werecollected and the solvent was evaporated, yielding 281 mg (66%) ofintermediate 58 as a yellow oil.

17) Preparation of Intermediate 59

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (200 mg, 0.0013 mol) and withaddition of triethylamine (0.2 ml). After workup, the residue waspurified by column chromatography over silica gel (40-63 μm) (eluent:EtOAc). The pure fractions were collected and the solvent wasevaporated, yielding 80 mg (25%) of intermediate 59 as a yellow oil.

18) Preparation of Intermediate 60

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (200 mg, 0.0013 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: DCM/MeOH 95/5). The pure fractions werecollected and the solvent was evaporated, yielding 176 mg (68%) ofintermediate 60 as a pale yellow oil.

19) Preparation of Intermediate 61

Similar procedure as method 6 (see Example A13) was followed, startingfrom 1-(4-chloro-2-pyridinyl)-ethanone (200 mg, 0.0013 mol). Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm)(eluent: EtOAc). The pure fractions were collected and thesolvent was evaporated, yielding 274 mg (77%) of intermediate 61 as ayellow oil.

20) Preparation of Intermediate 62

Method 3

A solution of 4-chloro-α-methyl-2-pyridinemethanol (200 mg, 0.0013 mol)in THF (3 ml) was added dropwise to a mixture of sodium hydride (60%weight in mineral oil) (56 mg, 0.0014 mol) in THF (1 ml) at 0° C. underargon. The mixture was heated up to 70° C. and stirred 3 hours, thencooled down to 0° C., and iodoethane (0.102 ml, 0.0013 mol) was addeddrop wise. The mixture was heated up to 70° C. for 2 hours, cooled downto 0° C., poured out into iced water, and extracted twice with DCM, andonce with EtOAc. The organic layer was separated, dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent:cyclohexane/EtOAc 90/10). The pure fractions were collected and thesolvent was evaporated, yielding 106 mg (45%) of intermediate 62 as abrown oil.

21) Preparation of Intermediate 63

Similar procedure as method 3 (see Example A22/20) was followed,starting from 4-chloro-α-methyl-2-pyridinemethanol (200 mg, 0.0013 mol).After workup, the residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: cyclohexane/EtOAc 90/10). The purefractions were collected and the solvent was evaporated, yielding 85 mg(39%) of intermediate 63 as a pale yellow oil.

22) Preparation of Intermediate 64

Method 4

4-Chloro-2-pyridinemethanol (400 mg, 0.0028 mol) was dissolved inchloroform (24 ml). Thionyl chloride (0.40 ml, 0.0056 mol) and DMF (2drops) were added. The mixture was stirred 4 hours at 80° C. The solventwas evaporated. The residue was taken back in MeOH (18 ml) andethanolamine (1.38 ml, 0.014 mol) was added. The mixture was stirred 4hours at 80° C. The solvent was evaporated. The residue was poured outonto water and extracted with EtOAc. The organic layer was separated,washed with a saturated solution of sodium hydrogen carbonate, dried(MgSO₄), filtered, and the solvent was evaporated. The residue waspurified by column chromatography over silica gel (40-63 μm) (eluent:DCM/MeOH 85/15). The pure fractions were collected and the solvent wasevaporated, yielding 310 mg (60%) of intermediate 64 as an orange oil.

23) Preparation of Intermediate 65

Similar procedure as method 4 (see Example A22/22) was followed,starting from 2-morpholin-4-yl-ethylamine (0.45 ml, 0.0034 mol) and4-chloro-2-pyridinemethanol (200 mg, 0.0014 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 39 mg (23%) of intermediate 65 as ayellow oil.

24) Preparation of Intermediate 66

Similar procedure as method 4 (see Example A22/22) was followed,starting from a 33% methyl amine solution in EtOH (10 ml) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH/NH₄OH 85/15/1). The pure fractions were collected andthe solvent was evaporated, yielding 130 mg (40%) of intermediate 66 asan orange oil.

25) Preparation of Intermediate 67

Similar procedure as method 4 (see Example A22/22) was followed,starting from a 2.0 M ethyl amine solution in THF (3.5 ml, 0.0069 mol)and 4-chloro-2-pyridinemethanol (200 mg, 0.0014 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 85/15). The pure fractions were collected and thesolvent was evaporated, yielding 45 mg (19%) of intermediate 67 as anorange oil.

26) Preparation of Intermediate 68

Similar procedure as method 4 (see Example A22/22) was followed,starting from 3-amino-propionic acid ethyl ester (2.45 g, 0.020 mol) and4-chloro-2-pyridinemethanol (600 mg, 0.0042 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 85/15). The pure fractions were collected and thesolvent was evaporated, yielding 730 mg (71%) of intermediate 68 as anorange liquid.

27) Preparation of Intermediate 69

Intermediate 68 (350 mg, 0.0015 mol) was dissolved in MeOH (5 ml) andcooled down at 0° C. Sodium borohydride (300 mg, 0.0078 mol) was slowlyadded. The mixture was stirred at 80° C. for 9 hours. The reaction wasquenched with water and the solvent was evaporated. The residue wasextracted with EtOAc. The organic layer was separated, washed with asaturated solution of sodium hydrogen carbonate, dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH85/15). The pure fractions were collected and the solvent wasevaporated, yielding 70 mg (23%) of intermediate 69 as a colorless oil.

28) Preparation of Intermediate 70

Similar procedure as method 4 (see Example A22/22) was followed,starting from amino-acetonitrile (1.2 g, 0.013 mol) and4-chloro-2-pyridinemethanol (500 mg, 0.0034 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 160 mg (25%) of intermediate 70 as anorange liquid.

29) Preparation of Intermediate 71

Similar procedure as method 4 (see Example A22/22) was followed,starting from N-methylpiperazine (1.16 ml, 0.010 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 325 mg (69%) of intermediate 71 as ayellow oil.

30) Preparation of Intermediate 72

Similar procedure as method 4 (see Example A22/22) was followed,starting from diethylamine (1.45 ml, 0.014 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 300 mg (43%) of intermediate 72 as ayellow liquid.

31) Preparation of Intermediate 73

Similar procedure as method 4 (see Example A22/22) was followed,starting from 3-aminopropionitrile (1.02 ml, 0.014 mol) and4-chloro-2-pyridinemethanol (500 mg, 0.0034 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 180 mg (27%) of intermediate 73 as ayellow oil.

32) Preparation of Intermediate 74

Similar procedure as method 4 (see Example A22/22) was followed,starting from phenethylamine (0.52 ml, 0.0042 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc). The pure fractions were collected and the solvent wasevaporated, yielding 110 mg (22%) of intermediate 74 as a colorless oil.

33) Preparation of Intermediate 75

Similar procedure as method 4 (see Example A22/22) was followed,starting from 3-phenyl-propylamine (470 mg, 0.0035 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 85/15). The pure fractions were collected and thesolvent was evaporated, yielding 90 mg (17%) of intermediate 75 as anorange oil.

34) Preparation of Intermediate 76

Method 5

A mixture of 4-chloro-2-pyridinecarboxaldehyde (200 mg, 0.0014 mol),N-(3-aminopropyl)morpholine (224 mg, 0.0015 mol), para-toluene sulfonicacid (134 mg, 0.00070 mol) and 3 Å molecular sieves was stirred at roomtemperature under Argon for 7 hours. Molecular sieves were filtered off,the reaction mixture was cooled down to 0° C., and sodium borohydride(107 mg, 0.0028 mol) was slowly added. The mixture was stirred at roomtemperature for 17 hours, poured out into water and extracted with DCM.The organic layer was separated, washed with a saturated solution ofhydrogen carbonate, dried (MgSO₄), filtered, and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: DCM/MeOH/NH₃ 85/15/3). The pure fractionswere collected and the solvent was evaporated, yielding 230 mg (60%) ofintermediate 76 as a yellow oil.

35) Preparation of Intermediate 77

Similar procedure as method 4 (see Example A22/22) was followed,starting from benzylamine (0.46 ml, 0.0042 mol) and4-chloro-2-pyridinemethanol (300 mg, 0.0021 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc/cyclohexane 50/50). The pure fractions were collected andthe solvent was evaporated, yielding 240 mg (50%) of intermediate 77 asa colorless oil.

36) Preparation of Intermediate 78

Similar procedure as method 3 (see Example A22/20) was followed,starting from bromoethyl methyl ether (0.13 ml, 0.0014 mol) and4-chloro-2-pyridinemethanol (200 mg, 0.0014 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: AcOEt/cyclohexene 30/70). The pure fractions were collected andthe solvent was evaporated, yielding 67 mg (24%) of intermediate 78 as ayellow oil.

37) Preparation of Intermediate 79

Similar procedure as method 4 (see Example A22/22) was followed,starting from 4-phenyl-butylamine (0.55 ml, 0.0035 mol) and4-chloro-2-pyridinemethanol (250 mg, 0.0017 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 260 mg (55%) of intermediate 79 as ayellow oil.

38) Preparation of Intermediate 80

Similar procedure as method 3 (see Example A22/20) was followed,starting from (3-bromo-propyl)-benzene (0.27 ml, 0.0018 mol) and4-chloro-2-pyridinemethanol (200 mg, 0.0014 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: AcOEt/cyclohexane 10/90). The pure fractions were collected andthe solvent was evaporated, yielding 57 mg (16%) of intermediate 80 ascolorless oil.

39) Preparation of Intermediate 81

Similar procedure as method 3 (cA22/20) was followed, starting from1-Bromo-2-ethoxy-ethane (589 mg, 0.0052 mol) and4-chloro-2-pyridinemethanol (500 mg, 0.0034 mol). After workup, theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc/cyclohexane 10/90). The pure fractions were collected andthe solvent was evaporated, yielding 270 mg (36%) of intermediate 81 asa colorless oil.

40) Preparation of Intermediate 82

Similar procedure as for method 1 (sec Example A22/10) was followed,starting from 4-chloro-2-pyridinecarboxaldehyde (500 mg, 0.0035 mol).After workup, the residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The purefractions were collected and the solvent was evaporated, yielding 136 mg(22%) of intermediate 82 as a yellow oil.

Example A23 a) Preparation of Intermediate 83

37% Hydrochloric acid solution (14.3 ml) was added to a solution of2-ethoxy-4-nitro-benzenamine (10.5 g, 0.0577 mol) in acetic acid (210ml) and the mixture was stirred at room temperature for 30 minutes. Thena solution of sodium nitrite (4.4 g, 0.0635 mol) in water (15 ml) wasadded drop wise and the mixture was stirred at 0° C. for 30 minutes. Acooled solution of potassium iodide (19.2 g, 0.1157 mol) and iodine (7.3g, 0.0288 mol) in water (70 ml) was added drop wise at 0° C. The mixturewas stirred 30 minutes at 0° C. and 16 hours at room temperature. Theresulting precipitate was filtered off, washed with water and thendissolved in DCM. The organic solution was washed with a saturatedsolution of sodium hydrogen carbonate, dried (MgSO₄), filtered and thesolvent was evaporated, yielding 13.7 g (81%) of intermediate 83 as ayellow solid.

b) Preparation of Intermediate 84

A mixture of intermediate 83 (700 mg, 0.0024 mol), 6-methoxytryptamine(505 mg, 0.0026 mol),dichloro[1,1′-bis(diphenylphosphino)ferrocene]palladium (II)dichloromethane adduct (78 mg, 0.00011 mol),1,1′Bis(diphenylphosphino)ferrocene (177 mg, 0.00032 mol) and sodiumtert-butoxide (255 mg, 0.0026 mol) in THF (95 ml) was heated at 100° C.for 3 hours and at 120° C. for 1.5 hour. After filtration through acelite pad, the solvent was evaporated and the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM). The purefractions were collected and the solvent was evaporated, yielding 464 mg(55%) of intermediate 84 as a yellow solid.

c Preparation of Intermediate 85

A mixture of intermediate 84 (see Example A23/b) (773 mg, 0.0022 mol)and Raney Nickel (50% slurry in water) in ethanol (8.5 ml) and THF (6.8ml) was stirred at room temperature under 1 atmosphere of hydrogen for24 hours. After filtration through celite, the solvent was evaporated,yielding 697 mg (98%) of intermediate 85 as a violet foam.

Example A24 Preparation of Intermediates 86 and 87

A mixture of 3,4,5-trichloro-pyridazine (200 mg, 0.0011 mol),intermediate 2 (see Example A1/b) (273 mg, 0.0011 mol) anddiisopropylamine (0.38 ml, 0.0011 mol) was stirred in 2-propanol (4.0ml) at 80° C. for 1 hour. The solvent was evaporated, and the crudemixture was taken back in EtOAc. The organic layer was washed with asaturated solution of sodium bicarbonate and with brine, dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent:EtOAc/cyclohexane 50/50). The pure fractions were collected and thesolvent was evaporated, yielding 179 mg (41%) of intermediate 86 andintermediate 87 as a 1/1 mixture of two pyridazine compounds.

Example A25 a) Preparation of Intermediate 88

A mixture of 4-oxiranyl-1-(phenylmethyl)-piperidine (0.069 mol) in MeOH(300 ml) and NaOCH₃ (0.069 mol) was stirred and refluxed for 6 hours.The solvent was evaporated, then the residue was taken up in water andextracted with DCM. The organic layer was separated, dried, filtered andthe solvent was evaporated. The residue was purified by columnchromatography over silica gel (eluent: DCM/MeOH 98/2, 90/10, 85/15).The product fractions were collected and the solvent was evaporated,yielding 5.0 g (29%) of intermediate 88.

b) Preparation of Intermediate 89

A mixture of intermediate 88 (see Example A25/a) (0.02 mol) in MeOH (100ml) was hydrogenated with Pd/C 10% (1 g) as a catalyst. After uptake ofH₂ (1 equiv.), the catalyst was filtered off and the filtrate wasevaporated, yield 3.18 g (100%) of intermediate 89.

Example A26 a) Preparation of Intermediate 90

Similar procedure as for method 5 (see Example A22/34) was followed,starting from benzyl N-(2-aminoethyl)carbamate hydrochloride (475 ml,0.0020 mol) and 4-chloro-2-pyridinecarboxaldehyde (265 mg, 0.0019 mol)and with addition of triethylamine (0.29 ml, 0.0021 mol). After workup,the residue was purified by column chromatography over silica gel (40-63μm) (eluent: DCM/MeOH 95/5). The pure fractions were collected and thesolvent was evaporated, yielding 150 mg (25%) of intermediate 90 as acolorless oil.

B. Preparation of the Final Compounds Example B1 Preparation of Compound1

A mixture of 4-chloro-quinoline (0.0009 mol) and intermediate 2 (0.001mol) in 2-propanol (5 ml) was stirred and refluxed for 6 hours, thenbrought to room temperature. The solvent was evaporated. The residue wasbasified with potassium carbonate 10% and extracted with DCM. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue (0.38 g) was purified by columnchromatography over silica gel (10 μm) (eluent: DCM/MeOH/NH₄OH97/3/0.5). The pure fractions were collected and the solvent wasevaporated. 2-Propanol and HC 1/2-propanol were added. The mixture wasstirred for 30 minutes, then brought to room temperature. Theprecipitate was filtered off and dried with diethyl ether, yielding 0.09g (23%) of compound 1, melting point 170° C.

Example B2 Preparation of Compound 2

A mixture of 4-bromo-pyridine, hydrochloride (0.0044 mol) andintermediate 4 (0.0044 mol) in acetic acid (13 ml) was stirred at 110°C. for 45 minutes, then cooled to room temperature, poured out into icewater, basified with potassium carbonate and extracted with DCM. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue (1.4 g) was purified by columnchromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH93/7/0.5). The pure fractions were collected and the solvent wasevaporated. The residue (0.38 g) was dissolved in 2-propanol/diethylether and converted into the hydrochloric acid salt. The precipitate wasfiltered off and dried, yielding 0.385 g (20%) of compound 2, meltingpoint 150° C.

Example B3 Preparation of Compound 3

A mixture of 4-chloro-2 (1H)-quinolinone (0.0011 mol) and intermediate 2(0.0016 mol) was stirred at 130° C. for 5 hours, then stirred at 160° C.overnight and brought to room temperature. The residue was purified bycolumn chromatography over silica gel (35-70 μm) (eluent: DCM/MeOH/NH₄OH95/5/0.1). The pure fractions were collected and the solvent wasevaporated. The residue (0.12 g) was taken up in acetonitrile. Theprecipitate was filtered off and dried, yielding 0.045 g (10%) ofcompound 3, melting point 238° C.

Example B4 Preparation of Compound 4

A mixture of 4-chloro-6,7-dihydro-5H-cyclopenta[b]pyridine (0.0006 mol)and intermediate 2 (0.0006 mol) in acetic acid (2 ml) was stirred at100° C. for 30 minutes and brought to room temperature. Water and thensodium hydroxide (3N) were added and the resulting mixture was extractedwith DCM. The organic layer was separated, dried (MgSO₄), filtered, andthe solvent was evaporated. The obtained residue (0.233 g) was purifiedby column chromatography over silica gel (10 μm) (DCM/MeOH/NH₄OH97/3/0.3). The pure fractions were collected and the solvent wasevaporated, yielding 0.025 g (11%) of compound 4.

Example B5 Preparation of Compound 5

A mixture of 4-chloro-5,6,7,8-tetrahydro-quinoline (0.0009 mol) andintermediate 2 (0.0009 mol) in DMF (3 ml) was stirred at 100° C. for 3hours and then brought to room temperature. The mixture was poured outinto ice water and sodium hydroxide (3N) and was then extracted withDCM. The organic layer was separated, dried (MgSO₄), filtered and thesolvent was evaporated. The obtained residue (0.49 g) was purified bycolumn chromatography over silica gel (5 μm) (DCM/MeOH/NH4OH 99/1/0.05to 80/20/0.5). The pure fractions were collected and the solvent wasevaporated, yielding 0.054 g (16%) of compound 5.

Example B6 Preparation of Compound 6

Lithium aluminum hydride (0.0032 mol) was added portionwise at 0° C. toa mixture of N-methoxy-N-methyl-1H-indole-3-acetamide (0.0032 mol) inTHF (5 ml) under N₂ flow. The mixture was stirred for 1 hour. Potassiumhydrogen sulfate (5%) was added. The mixture was extracted with diethylether. The organic layer was separated, dried (MgSO₄), filtered, and thesolvent was evaporated. This mixture has to be used immediately.Intermediate 7 (0.0016 mol), cyanoborohydride (0.0022 mol) on polymersupport (Amberlite IRA-300 BH₃CN form-capacity BH₃CN-=2.5/4.5 mmol/gresin) and acetic acid (few drops) in MeOH (5 ml) were added to themixture obtained. The mixture was stirred for 12 hours. The precipitatewas filtered off and dried. The residue was purified by columnchromatography over silica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH97/3/0.3). The pure fractions were collected and the solvent wasevaporated. The residue (0.14 g) was taken up in HC1/2-propanol. Theprecipitate was filtered off and dried, yielding 0.125 g (29%) ofcompound 6, melting point 160° C.

Example B7 Preparation of Compound 7

A mixture of N-4-pyridinyl-1,4-benzenediamine (0.0016 mol) and6-methoxy-1H-indole-3-carboxaldehyde (0.0016 mol) in MeOH (20 ml) wasstirred and refluxed overnight. Sodium tetrahydroborate (0.0016 mol) wasadded. The mixture was stirred at room temperature for 4 hours, pouredout on ice and extracted with DCM. The organic layer was separated,dried (MgSO₄), filtered, and the solvent was evaporated. The residue waspurified twice by column chromatography over kromasil (10 μm) (eluent:DCM/MeOH/NH₄OH 92/8/0.5 then toluene/2-propanol/NH₄OH 85/15/1). The purefractions were collected and the solvent was evaporated. The residue wascrystallized from acetonitrile. The precipitate was filtered off anddried, yielding 0.131 g (23%) of compound 7, melting point 145° C.

Example B8 Preparation of Compound 8

A mixture of 4-bromo-pyridine, hydrochloride (0.0069 mol) andintermediate 8 (0.008 mol) in acetic acid (7 ml) was stirred at 120° C.for 1 hour, then brought to room temperature. Water was added. Themixture was basified with potassium carbonate and extracted twice withDCM/MeOH (95/5). The organic layer was separated, dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified byflash column chromatography over silica gel (35-70 μm) (eluent:DCM/MeOH/NH₄OH 92/8/0.5). The pure fractions were collected and thesolvent was evaporated, yielding 1.6 g (65%) of compound 8, meltingpoint 208° C.

Example B9 Preparation of Compound 9

4-Pyridinecarboxaldehyde (0.0005 mol) then cyanoborohydride (0.0004 mol)on polymer support (Amberlite IRA-300 BH₃CN form-capacity BH₃CN-=2.5/4.5mmol/g resin then acetic acid (3 drops) were added to a mixture ofintermediate 2 (0.0004 mol) in MeOH (10 ml). The mixture was stirred atroom temperature for 3 hours. The precipitate was filtered off andwashed with MeOH. The filtrate was evaporated. The residue (0.17 g),which is a mixture of the targeted compound 9 and of the correspondingnot reduced intermediate imine, was dissolved in MeOH (20 ml). Sodiumtetrahydroborate (0.02 g) was added portionwise. The mixture was stirredfor 30 minutes. Water was added. MeOH was partly evaporated. The mixturewas extracted with EtOAc. The organic layer was washed with water, dried(MgSO₄), filtered and the solvent was evaporated. The residue (0.05 g)was purified by column chromatography over silica gel (10 μm) (eluent:DCM/MeOH/NH₄OH 98/2/0.4). The pure fractions were collected and thesolvent was evaporated. The residue (0.034 g) was dissolved in2-propanone and converted into the ethanedioic acid salt. Theprecipitate was filtered off and dried, yielding 0.036 g (16%) ofcompound 9, melting point 132° C.

Example B10 Preparation of Compound 10

A mixture of 4-bromo-pyridine, hydrochloride (0.001 mol) andintermediate 10 (0.0005 mol) in acetic acid (2 ml) was stirred at 120°C. for 1 hour, then brought to room temperature. Ice then sodiumhydroxide 3N were added. The mixture was extracted twice with DCM. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue was purified by column chromatography oversilica gel (10 μm) (eluent: DCM/MeOH/NH₄OH 96/4/0.5). The pure fractionswere collected and the solvent was evaporated. The residue (0.064 g,29%) was dissolved in 2-propanol/diethyl ether and converted into thehydrochloric acid salt. The precipitate was filtered off and dried,yielding 0.082 g (29%) of compound 10, melting point>250° C.

Example B11 Preparation of Compound 11

Lithium aluminum hydride (0.0145 mol) was added to a mixture ofintermediate 13 (0.0036 mol) in THF (100 ml). The mixture was stirredand refluxed for 3 hours, then brought to room temperature. EtOAc wasadded. A minimum of water was added. The mixture was filtered overcelite. Celite was washed with EtOAc. The organic layer was separated,dried (MgSO₄), filtered, and the solvent was evaporated. The residue(1.1 g) was purified by column chromatography over silica gel (15-40 μm)(eluent: DCM/MeOH/NH₄OH 93/7/0.5). The pure fractions were collected andthe solvent was evaporated. The residue (0.25 g) was crystallized fromacetonitrile/diethyl ether. The precipitate was filtered off and dried,yielding 0.11 g (12%) of compound 11, melting point 122° C.

Example B12 Preparation of Compound 86

A mixture of 4-chloro-2-pyridinecarbonitrile (154 mg, 0.0011 mol),intermediate 2 ((280 mg, 0.0011 mol) and a 5N hydrochloride solution in2-propanol (0.19 ml, 0.0011 mol) in DMF (2 ml) was stirred under argonat 100° C. for 24 hours, then cooled down to room temperature and pouredout into water. The resulting mixture was made alkaline with a saturatedsodium bicarbonate solution and extracted twice with EtOAc. The organiclayer was washed successively with a saturated solution of sodiumbicarbonate and with brine, dried (MgSO₄), filtered, and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: EtOAc/cyclohexane 50/50). The purefractions were collected and the solvent was evaporated, yielding 130 mg(33%) of compound 86 as a beige foam.

Example B13 Preparation of Compound 87

A mixture of compound 86 (110 mg, 0.00031 mol), sodium azide (22 mg,0.00034 mol) and zinc bromide (70 mg, 0.00031 mol) in water (1 ml) and2-propanol (0.25 ml) was stirred at 105° C. for 22 hours and was thencooled down to room temperature. A 0.25 N solution of sodium hydroxide(3 ml) was added and the mixture was stirred at room temperature for 1hour. The precipitate was filtered off, washed with MeOH, THF and1-butanol. The organic layer was evaporated and the resulting solid waswashed with MeOH and dried, yielding 26 mg (21%) of compound 87 as abeige solid, melting point 210° C.

Example B14 Preparation of Compound 88

Similar procedure as for compound 86 (method B12) was followed, startingfrom intermediate 14 (254 mg, 0.00076 mol) and intermediate 2 (191 mg,0.00076 mol). After workup, the residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH₄OH95/5/0.2). The pure fractions were collected and the solvent wasevaporated, yielding 139 mg (30%) of compound 88 as a grey foam.

Example B15 Preparation of Compound 89

A mixture of compound 88 (112 mg, 0.00020 mol) and palladium on carbon(10% wt) (43 mg, 0.000040 mol) in MeOH (1 ml) and EtOH (4 ml) wasstirred at room temperature under 1 atmosphere of hydrogen for 26 hours.After filtration through celite, the solvent was evaporated and theresidue was purified by SCX column chromatography. The pure fractionswere collected and the solvent was evaporated, yielding 27 mg (33%) ofcompound 89 as a grey foam.

Example B16 Preparation of Compound 90

Similar procedure as for compound 86 (method B12) was followed, startingfrom intermediate 15 (850 mg, 0.0024 mol) and intermediate 2 (561 mg,0.0022 mol), heating the mixture at 120° C. for 2 hours in a BiotageInitiator microwave apparatus. After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH95/5). The pure fractions were collected and the solvent was evaporated,yielding 680 mg (56%) of compound 90 as a brown foam.

Example B17 Preparation of Compound 91

Compound 90 (200 mg, 0.00036 mol) was dissolved in EtOH (10 ml) and MeOH(10 ml). Palladium on carbon (10% wt) (100 mg) was added. The mixturewas stirred at room temperature under hydrogen for 24 hours. The mixturewas filtrated on celite and washed with MeOH. The solvent wasevaporated, yielding 140 mg (92%) of compound 91 as a green oil.

Example B18 Preparation of Compound 92

Similar procedure as for compound 86 was followed, starting fromintermediate 90 (150 mg, 0.00047 mol) and intermediate 2 (107 mg,0.00042 mol), heating the mixture at 120° C. for 80 minutes in a BiotageInitiator microwave apparatus. After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH95/5). The pure fractions were collected and the solvent was evaporated,yielding 30 mg (14%) of intermediate 90 as a green oil.

Example B19 Preparation of Compound 93

Compound 92 (23 mg, 0.000043 mol) was dissolved in EtOH (1 ml) and MeOH(1 ml). Palladium on carbon (10% wt) (10 mg) was added. The mixture wasstirred at room temperature under hydrogen for 20 hours. The mixture wasfiltrated on celite and washed with MeOH. The solvent was evaporated.The residue was purified by SCX column chromatography, yielding 18 mg(100%) of compound 93 as a green oil.

Example B20 Preparation of Compound 94

Similar procedure as for compound 86 was followed, starting fromintermediate 17 (200 mg, 0.00056 mol) and intermediate 2 (142 mg,0.00056 mol), heating the mixture at 120° C. for 50 minutes in a BiotageInitiator microwave apparatus. After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH4OH85/15/1). The pure fractions were collected and the solvent wasevaporated, yielding 35 mg (11%) of compound 94 as a red oil.

Example B21 Preparation of Compound 95

Compound 94 (50 mg, 0.000088 mol) was dissolved in MeOH (3 ml). A 5Nhydrochloride solution in 2-propanol (5 ml) was added. The mixture wasstirred at room temperature for 17 hours. The solvent was evaporated.The residue was poured out onto water and extracted with EtOAc. Theorganic layer was separated, washed with a saturated solution of sodiumhydrogencarbonate, dried (MgSO4), filtered, and the solvent wasevaporated, yielding 15 mg (36%) of compound 95 as a yellow oil.

Example B22 Preparation of Compound 96

Similar procedure as for compound 86 was followed, starting fromintermediate 18 (160 mg, 0.00070 mol) and intermediate 2 (160 mg,0.00064 mol), heating the mixture at 120° C. for 1 hour in a BiotageInitiator microwave apparatus. After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH/NH4OH85/15/1). The pure fractions were collected and the solvent wasevaporated, yielding 100 mg (35%) of compound 96 as a green oil.

Example B23 Preparation of Compound 97

Compound 96 (50 mg, 0.00011 mol) was dissolved in THF (3 ml). Lithiumhydroxide (33 mg, 0.00079 mol) and water (1 drop) were added. Themixture was stirred at room temperature for 24 hours. The residue waspoured out onto water and extracted with EtOAc. The organic layer wasseparated, washed with a 4N sodium hydroxide solution.

The organic layer was separated, washed with a 3N hydrochloridesolution, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue was purified by SCX column chromatography, yielding 20 mg (41%)of compound 97 as a green oil.

Example B24 Preparation of Compound 98

Similar procedure as for compound 86 was followed, starting fromintermediate 19 (170 mg, 0.00079 mol) and intermediate 2 (180 mg,0.00071 mol), heating the mixture at 120° C. for 80 minutes in a BiotageInitiator microwave apparatus. After workup, the residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH85/15). The pure fractions were collected and the solvent wasevaporated, yielding 224 mg (73%) of compound 98 as a brown oil.

Example B25 Preparation of Compound 99

Compound 98 (100 mg, 0.00023 mol) was dissolved in MeOH (5 ml) andcooled at 0° C. Sodium borohydride (27 mg, 0.00069 mol) was addedslowly. The mixture was stirred at 80° C. for 4 hours. The reaction wasquenched with water and the solvent was evaporated. The residue wasextracted with EtOAc. The organic layer was separated, washed with asaturated solution of sodium hydrogencarbonate, dried (MgSO4), filtered,and the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 85/15). Thepure fractions were collected and the solvent was evaporated, yielding40 mg (43%) of compound 99 as a colorless oil.

Example B26 Preparation of Compound 100

A mixture of intermediate 20 (107 mg, 0.00047 mol), intermediate 2 (118mg, 0.00047 mol) and a 5N hydrochloride solution in 2-propanol (0.12 ml,0.00072 mol) in 1-methyl-2-pyrrolidinone (2.3 ml) was stirred underargon at 120° C. for 2 hours, then cooled down to room temperature andpoured out into water. The resulting mixture was basified with asaturated sodium hydrogen carbonate solution and extracted 3 times withEtOAc. The organic layer was isolated, washed with water and brine,dried (MgSO₄), filtered, and the solvent was evaporated. The residue waspurified by column chromatography over silica gel (40-63 μm) (eluent:DCM/MeOH/NH4OH 90/10/0.5). The pure fractions were collected and thesolvent was evaporated, yielding 61 mg (26%) of compound 100 as a beigefoam.

Example B27 Preparation of Compound 101

Lithium aluminium hydride (6.5 mg, 0.00017 mol) was added to a mixtureof intermediate 21 (53 mg, 0.00017 mol) in THF (1 ml) at 0° C. underargon. The mixture was stirred 1 hour at 0° C., quenched with a 5%solution of potassium hydrogen sulfate, and extracted with EtOAc. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue was solubilized in MeOH (1 ml) and addeddropwise to a mixture of N-4-pyridinyl-1,4-benzenediamine (32 mg,0.00017 mol), sodium cyanoborohydride (16 mg, 0.0025 mol), and aceticacid (1 drop) in MeOH (0.5 ml). The mixture was stirred 20 hours at roomtemperature, poured out onto water and extracted twice with EtOAc. Theorganic layer was separated, washed with a saturated solution of sodiumhydrogen carbonate and with brine, dried (MgSO₄), filtered, and thesolvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc/MeOH/NH₄OH90/10/0.3). The pure fractions were collected and the solvent wasevaporated, yielding 25 mg (34%) of compound 101 as a beige foam.

Example B28 Preparation of Compound 102

A mixture of intermediate 23 (500 mg, 0.0011 mol),2-chloro-4-bromopyridine (213 mg, 0.0011 mol),4,5-bis(diphenylphosphino)-9,9-dimethyxanthene (83 mg, 0.00014 mol),sodium tert-butoxide (264 mg, 0.0028 mol) in toluene (7.5 ml) wasdegassed under argon for 15 minutes.Tris(dibenzylideneacetone)dipalladium(0)-chloroform adduct (46 mg,0.000044 mol) was added. The mixture was heated at 100° C. for 90seconds in a Biotage Initiator microwave apparatus. The mixture wascooled down to room temperature then poured out into water and extractedtwice with EtOAc. The organic layer was washed twice with water, oncewith brine, dried (MgSO₄), filtered and the solvent was evaporated. Theresidue was purified by column chromatography over silica gel (40-63 μm)(eluent: EtOAc/cyclohexane 30/70). The pure fractions were collected andthe solvent was evaporated, yielding 254 mg (41%) of compound 102 as abeige foam.

Example B29 Preparation of Compound 103

Compound 102 (70 mg, 0.00012 mol) was dissolved in a 5N hydrochloridesolution in 2-propanol (1.5 ml). Water (2 drops) was added. The reactionmixture was stirred at room temperature for 5 hours. The reaction wasquenched and basified with a saturated sodium bicarbonate solution andextracted 3 times with EtOAc. The organic layer was dried (MgSO₄),filtered, and the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent:EtOAc/cyclohexane 50/50). The pure fractions were collected and thesolvent was evaporated, yielding 33 mg (76%) of compound 103 as a whitefoam.

Example B30 Preparation of Compound 104

A mixture of 4-chloro-α-methyl-2-pyridinemethanol (170 mg, 0.0011 mol)and intermediate 2 (271 mg, 0.0011 mol) in acetic acid (2 ml) wasstirred under Argon at 120° C. for 1 hour, then cooled down to roomtemperature and poured out into water. The resulting mixture wasbasified with a 4N sodium hydroxide solution and extracted twice withEtOAc. The organic layer was washed successively with a saturatedsolution of sodium bicarbonate and with brine, dried (MgSO₄), filtered,and the solvent was evaporated. The residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc/MeOH 80/20).The pure fractions were collected and the solvent was evaporated,yielding 190 mg (47%) of compound 104 as a beige foam.

Example B31 Preparation of Compound 105

A mixture of compound 104 (106 mg, 0.00029 mol) and activated manganeseoxide (148 mg, 0.0017 mol) in chloroform (4 ml) was stirred at roomtemperature for 6 hours. After filtration through a celite pad, thesolvent was evaporated and the residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc). The purefractions were collected and the solvent was evaporated, yielding 4 mg(3%) of compound 105 as an orange solid.

Example B32 Preparation of Compound 106

Acetamide oxime (11 mg, 0.00016 mol) was added at room temperature underargon to a mixture of activated 4 Å molecular sieves and sodium hydride(3.72 mg, 0.00016 mol) in THF (0.6 ml). The reaction mixture was stirredat 70° C. for 1.5 hour then cooled down to room temperature. A solutionof compound 200 (50 mg, 0.00013 mol) in THF (0.6 ml) was added. Thereaction mixture was stirred at 70° C. for 1 hour. The reaction wasquenched by addition of water and extracted twice with EtOAc. Theorganic layer was dried (MgSO₄), filtered, and the solvent wasevaporated. The residue was purified by column chromatography oversilica gel (40-63 μm) (eluent: EtOAc/cyclohexane 70/30). The purefractions were collected and the solvent was evaporated, yielding 16 mg(30%) of compound 106 as a yellow oil.

Example B33 Preparation of Compound 107

Similar procedure as for compound 106 was followed, starting frombenzamidoxime (38 mg, 0.00028 mol) and compound 200 (90 mg, 0.00023mol). After workup, the residue was purified by column chromatographyover silica gel (40-63 μm) (eluent: DCM/EtOAc 90/10). The pure fractionswere collected and the solvent was evaporated, yielding 13 mg (12%) ofcompound 107 as a yellow solid, melting point 170° C.-174° C.

Example B34 Preparation of Compound 108

Similar procedure as for compound 106 was followed, starting fromN′-Hydroxy-2-phenylethanimidamide (42 mg, 0.00028 mol) and compound 200(90 mg, 0.00023 mol). After workup, the residue was purified by columnchromatography over silica gel (40-63 μm) (eluent: EtOAc/cyclohexane50/50). The pure fractions were collected and the solvent wasevaporated, yielding 50 mg (45%) of compound 108 as a yellow solid,melting point 159° C.-161° C.

Example B35 Preparation of Compound 109

Similar procedure as for compound 86 was followed, starting from4-chloro-2,6-pyridinedicarboxylic acid, dimethyl ester (228 mg, 0.00099mol) and intermediate 2 (250 mg, 0.00099 mol), heating the mixture at120° C. for 2 hours in a Biotage Initiator microwave apparatus. Afterworkup, the residue was purified by column chromatography over silicagel (40-63 μm) (eluent: acetone/cyclohexane 30/70 to 60/40). The purefractions were collected and the solvent was evaporated, yielding 30 mg(7%) of compound 109 as a yellow foam.

Example B36 Preparation of Compound 110

A mixture of intermediate 27 (0.0016 mol) and intermediate 2 (0.0018mol) in acetic acid (35 ml) was stirred at 120° C. in a CEM Discovermicrowave oven (P=300 W) for 5 minutes, then brought back to roomtemperature. Ice and sodium hydroxide were added. The mixture wasfiltered over celite. Celite was washed with DCM/MeOH (95/5). Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue (1 g) was purified by column chromatographyover kromasil (15-40 μm) (eluent: DCM/MeOH/NH₄OH 95/5/0.5). The purefractions were collected and the solvent was evaporated. The residue(0.3 g) was crystallized from CH₃CN/MeOH. The precipitate was filteredoff and dried, yielding 0.182 g (29%) of compound 110, melting point136° C.

Example B37 Preparation of Compound 111

A mixture of intermediate 31 (0.0016 mol) and intermediate 2 (0.0018mol) in acetic acid (3.5 ml) was stirred in a CEM Discover microwaveoven (P=300 W) at 120° C. for 5 minutes, then cooled back to roomtemperature. Ice and concentrated NaOH were added. The mixture wasextracted twice with DCM. The organic layer was separated, dried(MgSO₄), filtered, and the solvent was evaporated. The residue (0.9 g)was purified by column chromatography over kromasil (10 μm) (eluent:DCM/MeOH/NH₄OH 93/7/0.5). The pure fractions were collected and thesolvent was evaporated. The residue (0.2 g) was crystallized fromCH₃CN/MeOH/acetone. The precipitate was filtered off and dried, yielding0.137 g (21%) of compound 111, melting point 104° C.

Example B38 Preparation of Compound 112

A mixture of intermediate 33 (0.0025 mol) and intermediate 27 (0.0025mol) in acetic acid (2.7 ml) was stirred in a CEM Discover microwaveoven (P=300 W) at 118° C. for 10 minutes, then brought to roomtemperature. Water and 3N sodium hydroxide were added. The mixture wasextracted with DCM. The organic layer was separated, dried (MgSO₄),filtered and the solvent was evaporated. The residue (1.42 g) waspurified by column chromatography over silica gel (10 μm) (eluent:DCM/MeOH/NH₄OH 95/5/0.5). The pure fractions were collected and thesolvent was evaporated. The residue (0.56 g) was taken up in2-propanone/CH₃CN. The precipitate was filtered off and dried, yielding0.506 g (35%) of compound 112, melting point 194° C.

Example B39 Preparation of Compound 113

Lithium borohydride (0.0026 mol) then MeOH (1 ml) were added portionwise at 0° C. to a solution of intermediate 35 (0.0002 mol) in THF (15ml) under N₂ flow. The mixture was stirred at 0° C. for 4 hours. Lithiumborohydride (15 eq) was added. The mixture was stirred at roomtemperature overnight. Lithium borohydride (10 eq) was added. Themixture was stirred at room temperature for 4 hours and 30 minutes.Lithium borohydride (15 eq) was added. The mixture was stirred at roomtemperature for 24 hours and poured out into water. MeOH and THF wereevaporated. DCM was added. The mixture was filtered, yielding 0.01 g ofa first batch of crude product. The filtrate was extracted with DCM. Theorganic layer was separated, dried (MgSO₄), filtered and the solvent wasevaporated, yielding 0.035 g of a second batch of crude product. Bothfractions were purified by column chromatography over kromasil (5 μm)(eluent: DCM/MeOH/NH4OH 95/5/0.5 to 85/15/1.5). The pure fractions werecollected and the solvent was evaporated, yielding 0.056 g (28%) ofcompound 113, melting point 154° C.

Example B41 Preparation of Compound 36

A mixture of 4-bromo-pyridine, hydrochloride (0.034 mol) andintermediate 2 (0.0374 mol) in acetic acid (13 ml) was stirred at 110°C. for 40 minutes, then cooled to room temperature, poured out into icewater and basified with potassium carbonate. DCM was added. The mixturewas stirred for 30 minutes, then filtered over celite. The filtrate wasdecanted. Celite was taken up in DCM/MeOH (95/5). The mixture wasstirred for 30 minutes, then filtered. Both filtrates were combined,dried (MgSO₄), filtered and the solvent was evaporated. The residue(16.8 g) was purified by column chromatography over silica gel (20-45μm) (eluent: DCM/MeOH/NH₄OH 92/8/0.5). The pure fractions were collectedand the solvent was evaporated. The residue (4.2 g) was taken up in2-propanone. The precipitate was filtered off and dried, yielding 3.6 g(32%) of compound 36, melting point 236° C.

Example B42 Preparation of Compound 115

N-(2-hydroxyethyl)piperazine (0.0019 mol), EDC (0.0019 mol), HOBT(0.0019 mol) and triethylamine (0.0019 mol) were added to a solution ofintermediate 41 (0.0013 mol) in DCM/DMF 75/25 (20 ml). The mixture wasstirred at room temperature overnight. Potassium carbonate 10% wasadded. The mixture was extracted with DCM. The organic layer wasseparated, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue (0.32 g) was purified by column chromatography over kromasil (10μm) (eluent: DCM/MeOH/NH₄OH 90/10/1). The pure fractions were collectedand the solvent was evaporated, yielding 0.027 g (4%) of compound 115.

Example B43 Preparation of Compound 116

1-[2-(2-hydroxyethoxy)ethyl]piperazine (0.0019 mol), EDCI (0.0019 mol),HOBT (0.0019 mol), and triethylamine (0.0019 mol) were added to asolution of intermediate 41 (0.0013 mol) in DCM/DMF 75/25 (20 ml). Themixture was stirred at room temperature overnight. Potassium carbonate10% was added. The mixture was extracted with DCM. The organic layer wasseparated, dried (MgSO₄), filtered, and the solvent was evaporated. Theresidue (0.38 g) was purified by column chromatography over kromasil (10μm) (eluent: DCM/MeOH/NH₄OH 90/10/1). The pure fractions were collectedand the solvent was evaporated, yielding 0.108 g (16%) of compound 116.

Example B44 Preparation of Compound 117

A mixture of intermediate 2 (0.002 mol) and4-chloro-1H-pyrrolo[2,3-d]pyrimidine (0.002 mol) in acetic acid (5 ml)was stirred in a CEM Discover microwave oven at 140° C. for 15 minutes.Acetic acid was evaporated. The crude product was dissolved in DCM. Theorganic layer was separated, dried (MgSO₄), filtered and the solvent wasevaporated. The residue (1 g) was purified by column chromatography oversilica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH 93/7/0.5). The purefractions were collected and the solvent was evaporated. The residue(0.27 g) was crystallized from acetonitrile. The precipitate wasfiltered off and dried, yielding 0.233 g (31%) of compound 117, meltingpoint 211° C.

Example B45 Preparation of Compound 118

5-amino-1-pentanol (0.0019 mol), EDC (0.0019 mol), HOBT (0.0019 mol),and triethylamine (0.0019 mol) were added to a solution of intermediate41 (0.0013 mol) in DCM/DMF 75/25 (10 ml). The mixture was stirred atroom temperature overnight, potassium carbonate 10% was added. Themixture was extracted with DCM. The organic layer was separated, dried(MgSO₄), filtered, and the solvent was evaporated. The residue (0.38 g)was purified by column chromatography over kromasil (10 μm) (eluent:DCM/MeOH/NH₄OH 90/10/1 to 80/20/2). The pure fractions were collectedand the solvent was evaporated, yielding 0.128 g of compound 118.

Example B46 Preparation of Compound 119

A mixture of intermediates 86 and 87 (179 mg, 0.00045 mol) and 10%palladium over carbon (20 mg) in EtOH was stirred at room temperatureunder 1 atmosphere of hydrogen for 16 hours. After filtration trough acelite pad, the solvent was evaporated. The residue was purified bycolumn chromatography over silica gel (40-63 μm) (eluent: DCM/MeOH 9/1).The pure fractions were collected and the solvent was evaporated,yielding 70 mg (47%) of compound 119 as a yellow foam.

Example B47 Preparation of Compound 120

A mixture of intermediate 2 (0.050 mg, 0.000199 mol),4-quinolinecarboxaldehyde (31 mg, 0.000199 mol) in MeOH were stirred andrefluxed overnight, then cooled to room temperature. Sodium borohydridewas added portionwise and the mixture was stirred at room temperaturefor 1 hour, hydrolyzed with water, extracted with DCM, dried over MgSO₄and concentrated. The residue was purified by column chromatography overkromasil (10 μm) (eluent: DCM/MeOH/NH₄OH 90/10/1 to 80/20/2). The purefractions were collected and the solvent was evaporated, yielding 0.045g (45%) of compound 120.

Example B48 Preparation of Compound 121

A mixture of 3-pyridinecarboxaldehyde (0.0028 mol) and intermediate 2(0.0028 mol) in MeOH (20 ml) was stirred and refluxed overnight, thenbrought to room temperature. Sodium tetrahydroborate (0.0028 mol) wasadded portionwise. The mixture was stirred at room temperature for 5hours. Ice and water were added. The mixture was extracted with DCM. Theorganic layer was separated, dried (MgSO₄), filtered, and the solventwas evaporated. The residue was purified by column chromatography oversilica gel (15-40 μm) (eluent: DCM/MeOH/NH₄OH 97/3/0.2). The purefractions were collected and the solvent was evaporated. The residue(0.4 g) was taken up in HCl/isopropanol/diethyl ether. The precipitatewas filtered off and dried. Ice and water were added. The mixture wasbasified with sodium hydroxide 3N. The mixture was extracted with DCM.The residue (0.3 g) was purified by column chromatography over silicagel (15-40 μm) (eluent: toluene/isopropanol/NH₄OH 90/10/0.5). The purefractions were collected and the solvent was evaporated. The residue(0.24 g) was taken up in isopropanol/HCl/isopropanol/diethyl ether. Theprecipitate was filtered off and dried, yielding 0.23 g (20%) ofcompound 121, isolated as a hydrochloric acid salt, melting point 130°C.

Table F-1 lists the compounds that were prepared according to one of theabove Examples. The following abbreviations were used in the tables:.C₂HF₃O₂ stands for the trifluoroacetate salt, int. stands forintermediate, comp. stands for compound, .HCl stands for hydrochloricacid salt, mp. stands for melting point, ms. stands for MASS spectrum.

TABLE F-1

C. Pharmacological Example

U87MG cells are human glioblastoma cells with wild type p53. In thiscell line MDM2 tightly controls p53 expression.

The capacity of the compounds to preserve p53 in U87MG cells wasmeasured with the p53 enzyme linked immunosorbent assay. The p53 assayis a “sandwich” enzyme immunoassay employing two polyclonal antibodies.A polyclonal antibody, specific for the p53 protein, has beenimmobilized onto the surface of the plastic wells. Any p53 present inthe sample to be assayed will bind to the capture antibody. Thebiotinylated detector polyclonal antibody also recognizes p53 protein,and will bind to any p53, which has been retained by the captureantibody. The detector antibody, in turn, is bond by horseradishperoxidase-conjugated streptavidin. The horseradish peroxidase catalysesthe conversion of the chromogenic substrate o-phenylene diamine, theintensity of which is proportional to the amount of p53 protein bond tothe plate. The colored reaction product is quantified using aspectrophotometer. Quantitation is achieved by the construction of astandard curve using known concentrations of purified recombinant HIStagged p53 protein (see example C.1.).

Cellular activity of the compounds of formula (I) was determined onU87MG tumour cells using a colorimetric assay for cell toxicity orsurvival (see example C.2).

C.1. p53 ELISA

U87MG cells (ATCC) were cultivated in Dulbecco's minimal essentialmedium (DMEM) supplemented with 10% foetal calf serum (FCS), 2 mML-glutamine, 1 mM sodium pyruvate, 1.5 g/l sodium bicarbonate andgentamycin at 37° C. in a humidified incubator with 5% CO₂.

U87MG cells were seeded at 30.000 cells per well in a 96 well plate,cultured for 24 hours and treated with compound for 16 hours at 37° C.in a humidified incubator. After incubation, the cells were washed oncewith phosphate-buffered saline and 30 μl, per well, low salt RIPA buffer(20 mM tris, pH7.0, 0.5 mM EDTA, 1% Nonidet P40, 0.5% DOC, 0.05% SDS, 1mM PMSF, 1 μg/ml aprotinin and 0.5 μl leupeptin) was added. Plates wereplaced on ice for 30 minutes to complete the lysis. p53 protein wasdetected in de lysates by using the sandwich ELISA, described below.

High binding polystyrene EIA/RIA 96 well plates (Costar 9018) werecoated with the capture antibody pAb122 (Roche 1413147) at aconcentration of 2 μg/ml in coating buffer (0.1 M NaHCO₃ pH8.2), 50 μlper well. The antibody was allowed to adhere overnight at 4° C. Coatedplates were washed once with phosphate-buffered saline (PBS)/0.05% Tween20 and 300 μl of blocking buffer (PBS, 1% bovine serum albumins (BSA))was added, for an incubation period of 2 hours at room temperature.Dilutions of purified recombinant HIS tagged p53 protein, ranging from3-200 ng/ml, were made in blocking buffer and used as standards.

Plates were washed twice with PBS/0.05% Tween 20 and blocking buffer orstandards were added at 80 μl/well. To the standards, 20 μl of lysisbuffer was added. The samples were added to the other wells at 20 μllysate/well. After an overnight incubation at 4° C., plates were washedtwice with PBS/0.05% Tween 20. Aliquots of 100 μl secondary polyclonalantibody p53 (FL-393) (Tebubio, sc-6243) at a concentration of 1 μg/mlin blocking buffer were added to each well and allowed to adhere for 2hours at room temperature. Plates were washed three times with PBS/0.05%Tween 20. Detection antibody anti-rabbit HRP (sc-2004, Tebubio) at 0.04μg/ml in PBS/1% BSA was added and incubated for 1 hour at roomtemperature. Plates were washed three times with PBS/0.05% Tween 20 and100 μl of substrate buffer was added (substrate buffer was preparedshortly before use by adding 1 tablet of 10 mg o-phenylene diamine (OPD)from Sigma and 125 μl 3% H₂O₂ to 25 ml OPD buffer: 35 mM citric acid,132 mM Na₂HPO₄, pH5.6). After 5 to 10 minutes, colour reaction wasstopped by adding 50 μl stop buffer (1 M H₂SO₄) per well. The absorbanceat dual wavelengths of 450/655 nm was measured using a Biorad microplate reader and the results were then analyzed.

For each experiment, controls (containing no drug) and a blankincubation (containing no cells or drugs) were run in parallel. Theblank value was subtracted from all control and sample values. For eachsample, the value of p53 (in absorbance units) was expressed as thepercentage of the value for p53 present in the control. Percentagepreservation higher than 130% was defined as significant. Herein theeffects of test compounds are expressed as the lowest dose giving atleast 130% of the value for p53 present in the control (LAD) (see tableF-2).

C.2. Proliferation Assay

All compounds tested were dissolved in DMSO and further dilutions weremade in culture medium. Final DMSO concentrations never exceeded 0.1%(v/v) in cell proliferation assays. Controls contained U87MG cells andDMSO without compound and blanks contained DMSO but no cells.

U87MG cells were seeded in 96-well cell culture plates at 3000cells/well/100 μl. 24 hours later, medium was changed and compoundand/or solvent were added to a final volume of 200 μl. Following 4 daysof incubation, medium was replaced by 200 μl fresh medium and cellgrowth was assessed using a MTT-based assay. Therefore, 25 μl of the MTTsolution (0.5% MTT research grade from Serva in phosphate-bufferedsaline) was added to each well and the cells were further incubated for2 hours at 37° C. The medium was then carefully aspirated and the blueMTT-formazan product was dissolved by adding to each well 25 μl 0.1 Mglycin and 100 μl DMSO. The plates were shaken for another 10 min on amicro plate shaker before reading absorbance at 540 nm by a Biorad microplate reader.

Within an experiment, the results for each experimental condition arethe mean of 3 replicate wells. For initial screening purposes, compoundswere tested at a single fixed concentration of 10⁻⁵ M. For activecompounds, the experiments were repeated to establish fullconcentration-response curves. For each experiment, controls (containingno drug) and a blank incubation (containing no cells or drugs) were runin parallel. The blank value was subtracted from all control and samplevalues. For each sample, the mean value for cell growth (in absorbanceunits) was expressed as a percentage of the mean value for cell growthof the control. When appropriate, IC₅₀-values (concentration of thedrug, needed to reduce cell growth to 50% of the control) were computedusing probit analysis for graded data (Finney, D. J., Probit Analyses,2^(nd) Ed. Chapter 10, Graded Responses, Cambridge University Press,Cambridge 1962). Herein the effects of test compounds are expressed aspIC₅₀ (the negative log value of the IC₅₀-value) (see table F-2).

TABLE F-2 Table F-2 lists the results of the compounds that were testedaccording to example C.1 and C.2. cell p53-elisa proliferation Co No LADpIC₅₀ 1 3.0E−08 >8.0 2 3.0E−07 7.2 3 >1.0E−05 5.3 4 3.0E−08 8.0 53.0E−08 6 >1.0E−05 5.5 7 1.0E−05 5.7 8 >1.0E−05 5.3 9 >1.0E−0510 >1.0E−05 5.9 11 >1.0E−05 5.3 12 3.0E−07 7.9 13 1.0E−07 7.6 14 3.0E−077.4 15 >1.0E−05 7.3 16 >1.0E−05 7.4 17 >1.0E−05 6.2 18 1.0E−07 6.3 193.0E−07 6.7 20 3.0E−07 7.0 21 3.0E−08 8.0 22 1.0E−07 7.7 23 1.0E−06 6.424 1.0E−07 >8.0 25 >1.0E−05 7.4 26 3.0E−06 7.0 27 3.0E−06 7.128 >1.0E−05 6.7 29 3.0E−06 6.6 30 >1.0E−05 6.5 31 >1.0E−05 5.9 323.0E−06 6.8 33 >1.0E−05 7.2 34 >1.0E−05 7.3 35 1.0E−06 7.4 36 1.0E−066.7 37 3.0E−07 6.8 38 >1.0E−05 39 1.0E−05 6.2 40 >1.0E−05 41 >1.0E−0542 >1.0E−05 43 >1.0E−05 44 >1.0E−05 45 >1.0E−05 6.0 46 1.0E−06 6.6 471.0E−05 6.8 48 1.0E−05 6.8 49 >1.0E−05 <5.0 50 3.0E−06 7.0 51 >1.0E−056.5 52 >1.0E−05 6.3 53 >1.0E−05 6.2 54 1.0E−06 6.9 55 3.0E−07 6.356 >1.0E−05 5.6 57 >1.0E−05 6.1 58 >1.0E−05 <5.0 59 1.0E−06 6.460 >1.0E−05 7.0 61 >1.0E−05 6.5 62 >1.0E−05 5.6 63 >1.0E−05 5.8 641.0E−06 6.4 65 >1.0E−05 <5.0 66 3.0E−07 7.2 67 >1.0E−05 5.9 68 >1.0E−055.6 69 1.0E−07 7.0 70 >1.0E−05 6.6 71 >1.0E−05 6.1 72 >1.0E−05 5.773 >1.0E−05 6.3 74 >1.0E−05 5.8 75 >1.0E−05 5.5 76 >1.0E−05 <5.077 >1.0E−05 5.5 78 >1.0E−05 5.0 79 >1.0E−05 5.6 82 >1.0E−05 <5.083 >1.0E−05 5.5 84 >1.0E−05 5.8 85 >1.0E−05 6.8 86 >1.00E−05 <5.087 >1.00E−05 <5.0 88 >1.00E−05 5.5 89 3.00E−06 5.4 90 >1.00E−05 5.691 >1.00E−05 5.6 92 >1.00E−05 5.5 93 >1.00E−05 <5.0 95 >1.00E−05 5.196 >1.00E−05 <5.0 97 1.00E−06 <5.0 98 >1.00E−05 5.4 99 1.00E−05 5.6100 >1.00E−05 5.4 101 >1.00E−05 5.6 102 >1.00E−05 <5.0 103 1.00E−05 5.4104 3.00E−06 5.5 105 >1.00E−05 5.1 106 >1.00E−05 5.8 107 >1.00E−05108 >1.00E−05 109 1.00E−06 <5.0 110 1.00E−07 8.0 111 1.00E−07 7.1 1121.00E−07 7.5 113 >1.00E−05 <5.0 114 >1.00E−05 <5.0 114 >1.00E−05 <5.0115 >1.00E−05 <5.0 116 >1.00E−05 <5.0 117 >1.00E−05 <5.0 118 >1.00E−05<5.0 119 >1.00E−05 <5.0 120 >1.00E−05 5.3 121 >1.00E−05 <5.0 123 5.3 1245.3 125 >1.00E−05 5.4 126 >1.00E−05 <5.0 127 >1.00E−05 5.1 128 >1.00E−055.5 129 3.00E−06 5.7 130 >1.00E−05 5.8 131 >1.00E−05 5.6 132 >1.00E−05<5.0 134 >1.00E−05 5.9 135 1.00E−06 6.0 136 >1.00E−05 5.7 137 >1.00E−055.5 138 >1.00E−05 5.8 139 >1.00E−05 5.7 140 >1.00E−05 5.6 141 1.00E−055.4 142 3.00E−06 5.5 143 >1.00E−05 5.5 144 5.5 145 5.6 146 >1.00E−05 5.1147 >1.00E−05 5.3 148 3.00E−07 5.5 149 >1.00E−05 5.7 150 1.00E−06 5.5151 1.00E−06 <5.0 152 >1.00E−05 5.0 153 >1.00E−05 5.6 154 >1.00E−05 5.5155 >1.00E−05 5.7 156 >3.00E−06 5.5 157 >1.00E−05 5.8 158 >1.00E−05 <5.0159 >1.00E−05 5.5 160 >1.00E−05 6.4 161 >1.00E−05 6.0 162 >1.00E−05 5.5163 >1.00E−05 5.5 164 >1.00E−05 5.6 165 3.00E−06 5.3 166 >1.00E−05 <5.0167 >1.00E−05 5.4 168 >1.00E−05 5.7 169 >1.00E−05 6.4 170 3.00E−07 5.5171 >1.00E−05 <5.0 172 >1.00E−05 <5.0 173 >1.00E−05 <5.0 174 >1.00E−05175 >1.00E−05 176 3.00E−06 177 3.00E−07 7.3 178 >1.00E−05 5.8 1793.00E−06 6.6 180 >1.00E−05 6.2 181 3.00E−07 6.6 182 >1.00E−05 5.8 1831.00E−05 6.3 184 >1.00E−05 6.0 185 3.00E−06 5.7 186 1.00E−06 6.0 1871.00E−06 6.4 188 1.00E−06 6.1 189 >1.00E−05 5.5 190 >1.00E−05 5.4191 >1.00E−05 5.5 192 1.00E−06 <5.0 193 3.00E−06 6.0 194 >1.00E−05 5.2195 1.00E−06 7.1 196 >1.00E−05 6.7 197 1.00E−07 6.6 198 1.00E−06 5.9199 >1.00E−05 5.7 201 >1.00E−05 5.5 202 >1.00E−05 5.5 203 >1.00E−05 5.5204 >1.00E−05 5.1 205 3.00E−06 6.1 206 >1.00E−05 5.5 207 >1.00E−05 6.1208 3.00E−06 <5.0 209 >1.00E−05 <5.0 210 >1.00E−05 <5.0 211 3.00E−07 7.2212 >1.00E−05 5.8 213 3.00E−06 <5.0 214 1.00E−06 <5.0 215 >1.00E−05 5.5216 1.00E−06 5.6 217 >1.00E−05 5.4 218 >1.00E−05 <5.0 219 >1.00E−05 5.4220 3.00E−06 5.2 221 >1.00E−05 5.4 222 >1.00E−05 5.4 223 >1.00E−05 6.1224 >1.00E−05 5.4 225 >1.00E−05 6.8 226 3.00E−06 5.5 227 >1.00E−05 5.1228 1.00E−06 5.1 229 1.00E−07 7.0

D. Composition Example: Film-coated Tablets

Preparation of Tablet Core

A mixture of 100 g of a compound of formula (I), 570 g lactose and 200 gstarch is mixed well and thereafter humidified with a solution of 5 gsodium dodecyl sulphate and 10 g polyvinyl-pyrrolidone in about 200 mlof water. The wet powder mixture is sieved, dried and sieved again. Thenthere is added 100 g microcrystalline cellulose and 15 g hydrogenatedvegetable oil. The whole is mixed well and compressed into tablets,giving 10.000 tablets, each comprising 10 mg of a compound of formula(I).

Coating

To a solution of 10 g methyl cellulose in 75 ml of denatured ethanolthere is added a solution of 5 g of ethyl cellulose in 150 ml ofdichloromethane. Then there are added 75 ml of dichloromethane and 2.5ml 1,2,3-propanetriol 10 g of polyethylene glycol is molten anddissolved in 75 ml of dichloromethane. The latter solution is added tothe former and then there are added 2.5 g of magnesium octadecanoate, 5g of polyvinyl-pyrrolidone and 30 ml of concentrated colour suspensionand the whole is homogenated. The tablet cores are coated with the thusobtained mixture in a coating apparatus.

The invention claimed is:
 1. A process of preparing a pharmaceuticalcomposition comprising pharmaceutically acceptable carriers and as anactive ingredient a therapeutically effective amount of a compound offormula (I),

a N-oxide form, an addition salt or a stereochemically isomeric formthereof, wherein m is 0, 1, or 2 and when m is 0 then a direct bond isintended; n is 0, 1, 2, or 3 and when n is 0 then a direct bond isintended; p is 0, or 1 and when p is 0 then a direct bond is intended; sis 0, or 1 and when s is 0 then a direct bond is intended; t is 0 or 1and when t is 0 then a direct bond is intended; X is C(═O) or CHR⁸;wherein R⁸ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, —C(═O)—NR¹⁷R¹⁸,hydroxycarbonyl, arylC₁₋₆alkyloxycarbonyl, heteroaryl,heteroarylcarbonyl, heteroarylC₁₋₆alkyloxycarbonyl, piperazinylcarbonyl,pyrrolidinyl, piperidinylcarbonyl, C₁₋₆alkyloxycarbonyl, C₁₋₆alkylsubstituted with a substituent selected from hydroxy, amino, aryl, andheteroaryl; C₃₋₇cycloalkyl substituted with a substituent selected fromhydroxy, amino, aryl, and heteroaryl; piperazinylcarbonyl substitutedwith hydroxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl;pyrrolidinyl substituted with hydroxyC₁₋₆alkyl; or piperidinylcarbonylsubstituted with one or two substituents selected from hydroxy,C₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyl(dihydroxy)C₁₋₆alkyl or C₁₋₆alkyloxy(hydroxy)C₁₋₆alkyl; R¹⁷ andR¹⁸ are each independently selected from hydrogen, C₁₋₆alkyl,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, arylC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyl(C₁₋₆alkyl) orhydroxyC₁₋₆alkyl(arylC₁₋₆alkyl);

 is —CR⁹═C< and then the dotted line is a bond, —C(═O)—CH<, —C(═O)—N<,—CHR⁹—CH< or —CHR⁹—N<; wherein each R⁹ is independently hydrogen orC₁₋₆alkyl; R¹ is hydrogen, aryl, heteroaryl, C₁₋₆alkyloxycarbonyl,C₁₋₁₂alkyl, or C₁₋₁₂alkyl substituted with one or two substituentsindependently selected from hydroxy, aryl, heteroaryl, amino,C₁₋₆alkyloxy, mono- or di(C₁₋₆alkyl)amino, morpholinyl, piperidinyl,pyrrolidinyl, piperazinyl, C₁₋₆alkylpiperazinyl,arylC₁₋₆alkylpiperazinyl, heteroarylC₁₋₆alkylpiperazinyl,C₃₋₇cycloalkylpiperazinyl and C₃₋₇cycloalkylC₁₋₆alkylpiperazinyl; R² ishydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy,heteroarylC₁₋₆alkyloxy, phenylthio, hydroxyC₁₋₆alkylcarbonyl, C₁₋₆alkylsubstituted with a substituent selected from amino, aryl and heteroaryl;or C₃₋₇cycloalkyl substituted with a substituent selected from amino,aryl and heteroaryl; R³ is hydrogen, C₁₋₆alkyl, heteroaryl,C₃₋₇cycloalkyl, C₁₋₆alkyl substituted with a substituent selected fromhydroxy, amino, aryl and heteroaryl; or C₃₋₇cycloalkyl substituted witha substituent selected from hydroxy, amino, aryl and heteroaryl; R⁴ andR⁵ are each independently hydrogen, halo, C₁₋₆alkyl, polyhaloC₁₋₆alkyl,cyano, cyanoC₁₋₆alkyl, hydroxy, amino or C₁₋₆alkyloxy; or R⁴ and R⁵together can optionally form a bivalent radical selected frommethylenedioxy or ethylenedioxy; R⁶ is hydrogen, C₁₋₆alkyloxycarbonyl orC₁₋₆alkyl; when p is 1 then R⁷ is hydrogen, arylC₁₋₆alkyl, hydroxy orheteroarylC₁₋₆alkyl; Z is a radical selected from

wherein each R¹⁰ or R¹¹ are each independently selected from hydrogen,halo, hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano,cyanoC₁₋₆alkyl, tetrazoloC₁₋₆alkyl, aryl, heteroaryl,heteroarylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl,heteroaryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, heteroarylcarbonyl,C₁₋₆alkylcarbonyl, arylC₁₋₆alkylcarbonyl, heteroarylC₁₋₆alkylcarbonyl,C₁₋₆alkyloxy, C₃₋₇cycloalkylcarbonyl, C₃₋₇cycloalkyl(hydroxy)C₁₋₆alkyl,arylC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonylC₁₋₆alkyloxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonylC₂₋₆alkenyl C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonyl, C₁₋₆alkylcarbonyloxy, aminocarbonyl,hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl,hydroxycarbonylC₁₋₆alkyl and —(CH₂)_(v)—(C(═O)_(r))—(CHR¹⁹)_(u)—NR¹³R¹⁴;wherein v is 0, 1, 2, 3, 4, 5, or 6 and when v is 0 then a direct bondis intended; r is 0, or 1 and when r is 0 then a direct bond isintended; u is 0 1, 2, 3, 4, 5 or 6 and when u is 0 then a direct bondis intended; R¹⁹ is hydrogen or C₁₋₆alkyl; R¹² is hydrogen, C₁₋₆alkyl,C₃₋₇cycloalkyl, C₁₋₆alkyl substituted with a substituent selected fromhydroxy, amino, C₁₋₆alkyloxy and aryl; or C₃₋₇cycloalkyl substitutedwith a substituent selected from hydroxy, amino, aryl and C₁₋₆alkyloxy;R¹³ and R¹⁴ are each independently selected from hydrogen, C₁₋₁₂alkyl,C₁₋₆alkylcarbonyl, C₁₋₆alkylsulfonyl, arylC₁₋₆alkylcarbonyl,C₃₋₇cycloalkyl, C₃₋₇cycloalkylcarbonyl, —(CH₂)_(k)—NR¹⁵R¹⁶, C₁₋₁₂alkylsubstituted with a substituent selected from hydroxy, hydroxycarbonyl,cyano, C₁₋₆alkyloxycarbonyl, C₁₋₆alkyloxy, aryl or heteroaryl; orC₃₋₇cycloalkyl substituted with a substituent selected from hydroxy,C₁₋₆alkyloxy, aryl, amino, arylC₁₋₆alkyl, heteroaryl orheteroarylC₁₋₆alkyl; or R¹³ and R¹⁴ together with the nitrogen to whichthey are attached can optionally form a morpholinyl, piperidinyl,pyrrolidinyl, piperazinyl, or piperazinyl substituted with a substituentselected from C₁₋₆alkyl, arylC₁₋₆alkyl, arylC₁₋₆alkyloxycarbonyl,heteroarylC₁₋₆alkyl, C₃₋₇cycloalkyl and C₃₋₇cycloalkylC₁₋₆alkyl; whereink is 0, 1, 2, 3, 4, 5, or 6 and when k is 0 then a direct bond isintended;  R¹⁵ and R¹⁶ are each independently selected from hydrogen,C₁₋₆alkyl, arylC₁₋₆alkyloxycarbonyl, C₃₋₇cycloalkyl, C₁₋₁₂alkylsubstituted with a substituent selected from hydroxy, C₁₋₆alkyloxy,aryl, and heteroaryl; and C₃₋₇cycloalkyl substituted with a substituentselected from hydroxy, C₁₋₆alkyloxy, aryl, arylC₁₋₆alkyl, heteroaryl,and heteroarylC₁₋₆alkyl; or  R¹⁵ and R¹⁶ together with the nitrogen towhich they are attached can optionally form a morpholinyl, a piperazinylor a piperazinyl substituted with C₁₋₆alkyloxycarbonl; aryl is phenyl ornaphthalenyl; each phenyl or naphthalenyl can optionally be substitutedwith one, two or three substituents each independently selected fromhalo, hydroxy, C₁₋₆alkyl, amino, polyhaloC₁₋₆alkyl and C₁₋₆alkyloxy; andeach phenyl or naphthalenyl can optionally be substituted with abivalent radical selected from methylenedioxy and ethylenedioxy;heteroaryl is pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl,thienyl, oxadiazolyl, tetrazolyl, benzofuranyl or tetrahydrofuranyl;each pyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,oxadiazolyl, tetrazolyl, benzofuranyl, or tetrahydrofuranyl canoptionally be substituted with one, two or three substituents eachindependently selected from halo, hydroxy, C₁₋₆alkyl, amino,polyhaloC₁₋₆alkyl, aryl, arylC₁₋₆alkyl or C₁₋₆alkyloxy; and eachpyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,benzofuranyl, or tetrahydrofuranyl can optionally be substituted with abivalent radical selected from methylenedioxy or ethylenedioxy; with theproviso that when m is 1; the substituents on the phenyl ring other thanR² are in the meta position; s is 0; and t is 0; then Z is a radicalselected from (a-1), (a-3), (a-4), (a-5), (a-6), (a-7), (a-8) or (a-9)wherein the pharmaceutically acceptable carriers and a compound offormula (I) are intimately mixed.
 2. A process for preparing a compoundof formula (I),

a N-oxide form, an addition salt or a stereochemically isomeric formthereof, wherein m is 0, 1, or 2 and when m is 0 then a direct bond isintended; n is 0, 1, 2, or 3 and when n is 0 then a direct bond isintended; p is 0, or 1 and when p is 0 then a direct bond is intended; sis 0, or 1 and when s is 0 then a direct bond is intended; t is 0 or 1and when t is 0 then a direct bond is intended; X is C(═O) or CHR⁸;wherein R⁸ is hydrogen, C₁₋₆alkyl, C₃₋₇cycloalkyl, —C(═O)—NR¹⁷R¹⁸,hydroxycarbonyl, arylC₁₋₆alkyloxycarbonyl, heteroaryl,heteroarylcarbonyl, heteroarylC₁₋₆alkyloxycarbonyl, piperazinylcarbonyl,pyrrolidinyl, piperidinylcarbonyl, C₁₋₆alkyloxycarbonyl, C₁₋₆alkylbstituted with a substituent selected from hydroxy, amino, aryl, andheteroaryl; C₃₋₇cycloalkyl substituted with a substituent selected fromhydroxy, amino, aryl, and heteroaryl; piperazinylcarbonyl substitutedwith hydroxy, hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl;pyrrolidinyl substituted with hydroxyC₁₋₆alkyl; or piperidinylcarbonylsubstituted with one or two substituents selected from hydroxy,C₁₋₆alkyl, hydroxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyl(dihydroxy)C₁₋₆alkyl or C₁₋₆alkyloxy(hydroxy)C₁₋₆alkyl; R¹⁷ andR¹⁸ are each independently selected from hydrogen, C₁₋₆alkyl,di(C₁₋₆alkyl)aminoC₁₋₆alkyl, arylC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyl,hydroxyC₁₋₆alkyl, hydroxyC₁₋₆alkyl(C₁₋₆alkyl) orhydroxyC₁₋₆alkyl(arylC₁₋₆alkyl);

 is —CR⁹═C< and then the dotted line is a bond, —C(═O)—CH<, —C(═O)—N<,—CHR⁹—CH< or —CHR⁹—N<; wherein each R⁹ is independently hydrogen orC₁₋₆alkyl; R¹ is hydrogen, aryl, heteroaryl, C₁₋₆alkyloxycarbonyl,C₁₋₆alkyl, or C₁₋₆alkyl substituted with one or two substituentsindependently selected from hydroxy, aryl, heteroaryl, amino,C₁₋₆alkyloxy, mono- or di(C₁₋₆alkyl)amino, morpholinyl, piperidinyl,pyrrolidinyl, piperazinyl, C₁₋₆alkylpiperazinyl,arylC₁₋₆alkylpiperazinyl, heteroarylC₁₋₆alkylpiperazinyl,C₃₋₇cycloalkylpiperazinyl and C₃₋₇cycloalkylC₁₋₆alkylpiperazinyl; R² ishydrogen, halo, C₁₋₆alkyl, C₁₋₆alkyloxy, arylC₁₋₆alkyloxy,heteroarylC₁₋₆alkyloxy, phenylthio, hydroxyC₁₋₆alkylcarbonyl, C₁₋₆alkylsubstituted with a substituent selected from amino, aryl and heteroaryl;or C₃₋₇cycloalkyl substituted with a substituent selected from amino,aryl and heteroaryl; R³ is hydrogen, C₁₋₆alkyl, heteroaryl,C_(3═)cvcloalkyl, C₁₋₆alkyl substituted with a substituent selected fromhydroxy, amino, aryl and heteroaryl; or C₃₋₇cycloalkyl substituted witha substituent selected from hydroxy, amino, aryl and heteroaryl; R⁴ andR⁵ are each independently hydrogen, halo, C₁₋₆alkyl, polyhaloC₁₋₆alkyl,cyano, cyanoC₁₋₆alkyl, hydroxy, amino or C₁₋₆alkyloxy; or R⁴ and R⁵together can optionally form a bivalent radical selected frommethylenedioxy or ethylenedioxy; R⁶ is hydrogen, C₁₋₆alkyloxycarbonyl orC₁₋₆alkyl; when p is 1 then R⁷ is hydrogen, arylC₁₋₆alkyl, hydroxy orheteroarylC₁₋₆alkyl; Z is a radical selected from

wherein each R¹⁰ or R¹¹ are each independently selected from hydrogen,halo, hydroxy, amino, C₁₋₆alkyl, nitro, polyhaloC₁₋₆alkyl, cyano,cyanoC₁₋₆alkyl, tetrazoloC₁₋₆alkyl, aryl, heteroaryl, arylC₁₋₆alkyl,heteroarylC₁₋₆alkyl, aryl(hydroxy)C₁₋₆alkyl,heteroaryl(hydroxy)C₁₋₆alkyl, arylcarbonyl, heteroarylcarbonyl,C₁₋₆alkylcarbonyl, arylC₁₋₆alkylcarbonyl, heteroarylC₁₋₆alkylcarbonyl,C₁₋₆alkyloxy, c₃₋₇cycloalkylcarbonyl, C₃₋₇cycloalkyl(hydroxy)C₁₋₆alkyl,arylC₁₋₆alkyloxyC₁₋₆alkyl, C₁₋₆alkyloxyC₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkylcarbonyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonylC₁₋₆alkyloxyC₁₋₆alkyl, hydroxyC₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonylC₂₋₆alkenyl C₁₋₆alkyloxyC₁₋₆alkyl,C₁₋₆alkyloxycarbonl, C₁₋₆alkylcarbonyloxy, aminocarbonyl,hydroxyC₁₋₆alkyl, aminoC₁₋₆alkyl, hydroxycarbonyl,hydroxycarbonylC₁₋₆alkyl and —(CH₂)_(v)—(C(═O)_(r))—(CHR¹⁹)_(u)—NR¹³R¹⁴;wherein v is 0, 1, 2, 3, 4, 5, or 6 and when v is 0 then a direct bondis intended; r is 0, or 1 and when r is 0 then a direct bond isintended; u is 0, 1, 2, 3, 4, 5, or 6 and when u is 0 then a direct bondis intended; R¹⁹ is hydrogen or C₁₋₆alkyl; R¹² is hydrogen, C₁₋₆alkyl,C₃₋₇cycloalkyl, C₁₋₆alkyl substituted with a substituent selected fromhydroxy, amino, C₁₋₆alkyloxy and aryl; or C₃₋₇cycloalkyl substitutedwith a substituent selected from hydroxy, amino, aryl and C₁₋₆alkyloxy;R¹³ and R¹⁴ are each independently selected from hydrogen, C₁₋₁₂alkyl,C₁₋₆alkylcarbonyl, C₁₋₆alkylsulfonyl, arylC₁₋₆alkylcarbonyl,C₃₋₇cycloalkyl, C₃₋₇cycloalkylcarbonyl, —(CH₂)—NR¹⁵R¹⁶, substituted witha substituent selected from hydroxy, hydroxycarbonyl, cyano,C₁₋₆alkyloxycarbonyl, C₁₋₆alkyloxy, aryl or heteroaryl; orC₃₋₇cycloalkyl substituted with a substituent selected from hydroxy,C₁₋₆alkyloxy, aryl, amino, arylC₁₋₆alkyl, heteroaryl orheteroarylC₁₋₆alkyl; or R¹³ and R¹⁴ together with the nitrogen to whichthey are attached can optionally form a morpholinyl, piperidinyl,pyrrolidinyl, piperazinyl, or piperazinyl substituted with a substituentselected from C₁₋₆alkyl, arylC₁₋₆alkyl, arylC₁₋₆alkyloxycarbonyl,heteroarylC₁₋₆alkyl, C₃₋₇cycloalkyl and C₃₋₇cycloalkylC₁₋₆alkyl; whereink is 0, 1, 2, 3, 4, 5, or 6 and when k is 0 then a direct bond isintended; R¹⁵ and R¹⁶ are each independently selected from hydrogen,C₁₋₆alkyl, arylC₁₋₆alkyloxycarbonyl, C₃₋₇cycloalkyl, C₁₋₁₂alkylsubstituted with a substituent selected from hydroxy, C₁₋₆alkyloxy,aryl, and heteroaryl; and C₃₋₇cycloalkyl substituted with a substituentselected from hydroxy, C₁₋₆alkyloxy, aryl, arylC₁₋₆alkyl, heteroaryl,and heteroarylC₁₋₆alkyl; or R¹⁵ and R¹⁶ together with the nitrogen towhich they are attached can optionally form a morpholinyl, a piperazinylor a piperazinyl substituted with C₁₋₆alkyloxycarbonyl; aryl is phenylor naphthalenyl; each phenyl or naphthalenyl can optionally besubstituted with one, two or three substituents each independentlyselected from halo, hydroxy, C₁₋₆alkyl, amino, polyhaloC₁₋₆alkyl andC₁₋₆alkyloxy; and each phenyl or naphthalenyl can optionally besubstituted with a bivalent radical selected from methylenedioxy andethylenedioxy; heteroaryl is pyridinyl, indolyl, quinolinyl, imidazolyl,furanyl, thienyl, oxadiazolyl, tetrazolyl, benzofuranyl ortetrahydrofuranyl; each pyridinyl, indolyl, quinolinyl, imidazolyl,furanyl, thienyl, oxadiazolyl, tetrazolyl, benzofuranyl, ortetrahydrofuranyl can optionally be substituted with one, two or threesubstituents each independently selected from halo, hydroxy, C₁₋₆alkyl,amino, polyhaloC₁₋₆alkyl, aryl, arylC₁₋₆alkyl or C₁₋₆alkyloxy; and eachpyridinyl, indolyl, quinolinyl, imidazolyl, furanyl, thienyl,benzofuranyl, or tetrahydrofuranyl can optionally be substituted with abivalent radical selected from methylenedioxy or ethylenedioxy; with theproviso that when m is 1; the substituents on the phenyl ring other thanR² are in the meta position; s is 0; and t is 0; then Z is a radicalselected from (a-1), (a-3), (a-4), (a-5), (a-6), (a-7), (a-8) or (a-9),characterized by a) reacting an intermediate of formula (II) with anintermediate of formula (III) wherein W is an appropriate leaving group,

b) converting a compound of formula (I) wherein X is C(═O), hereinreferred to as compounds of formula (I-b), into compounds of formula(I), wherein X is CH₂, herein referred to as compounds of formula (I-a),in the presence of lithium aluminium hydride in a suitable solvent,

c) reacting an appropriate carboxaldehyde of formula (IV), with anintermediate of formula (V), in the presence of an appropriate reagent,in a suitable solvent,

d) reacting an intermediate of formula (II) with an appropriatecarboxaldehyde of formula (VI) with the formation of a compound offormula (I), wherein t is 1, herein referred to as compounds of formula(I-c), or

e) reacting an intermediate of formula (VII) with lithium aluminiumhydride in a suitable solvent, with the formation of a compound offormula (I), wherein s is 1, herein referred to as compounds of formula(I-d),